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  • | BI-R2-DAS-Manuel utilisateur BI-B

    - 1 -

    DAS

    BI-R2-DAS-Manuel

    utilisateur BI-B

    2007-10-30 β-IMAGER User guide

    Référence Date

    ββββ IMAGER User guide

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    Table of Content

    1 IMAGER introduction ..................................................................................... 5

    1.11.11.11.1 IMAGERIMAGERIMAGERIMAGER description description description description................................................................................................................................................................................................................................................................................................................ 5555 1.21.21.21.2 IMAGERIMAGERIMAGERIMAGER operating principles operating principles operating principles operating principles............................................................................................................................................................................................................................................................ 8888 1.31.31.31.3 Control panelControl panelControl panelControl panel ............................................................................................................................................................................................................................................................................................................................................................ 9999 1.41.41.41.4 Operation precautionsOperation precautionsOperation precautionsOperation precautions....................................................................................................................................................................................................................................................................................................10101010

    1.4.1 ......Avoid the following situations:......................................................... 10 1.4.2 .....Moving the equipment..................................................................... 10 1.4.3 .....Gas exhaust .................................................................................... 10

    1.51.51.51.5 Normal operating conditionsNormal operating conditionsNormal operating conditionsNormal operating conditions................................................................................................................................................................................................................................................................ 11111111 1.5.1 ......Power supply....................................................................................11 1.5.2......Turning on the β IMAGER.................................................................11 1.5.3......Turning off the β IMAGER ................................................................11 1.5.4......Characteristics of the β IMAGER ......................................................11

    2 Sample handling ........................................................................................... 12 2.12.12.12.1 IntroductionIntroductionIntroductionIntroduction................................................................................................................................................................................................................................................................................................................................................................ 12121212 2.22.22.22.2 Glass slides cassetteGlass slides cassetteGlass slides cassetteGlass slides cassette............................................................................................................................................................................................................................................................................................................ 12121212 2.32.32.32.3 Tritium (TLC) cassetteTritium (TLC) cassetteTritium (TLC) cassetteTritium (TLC) cassette ........................................................................................................................................................................................................................................................................................................ 13131313

    2.3.1 .....Basic version:.................................................................................. 13 2.3.2 .....TLC version ...................................................................................... 13

    2.42.42.42.4 C14 (NonC14 (NonC14 (NonC14 (Non----tritiated membrane) cassettetritiated membrane) cassettetritiated membrane) cassettetritiated membrane) cassette.................................................................................................................................................................................................... 15151515 2.52.52.52.5 Sample Sample Sample Sample –––– Cassette matching Cassette matching Cassette matching Cassette matching............................................................................................................................................................................................................................................................ 17171717

    3 Acquisition Program: Presentation ................................................................ 18 3.13.13.13.1 IntroductionIntroductionIntroductionIntroduction................................................................................................................................................................................................................................................................................................................................................................18181818 3.23.23.23.2 MenusMenusMenusMenus................................................................................................................................................................................................................................................................................................................................................................................................ 19191919

    3.2.1 .....File Menu......................................................................................... 19 3.2.2 .....Tools Menu...................................................................................... 19 3.2.3 .....View Menu ......................................................................................20 3.2.4 .....Help Menu.......................................................................................20

    3.33.33.33.3 Shortcut IconsShortcut IconsShortcut IconsShortcut Icons ............................................................................................................................................................................................................................................................................................................................................ 20202020 3.43.43.43.4 Program WindowsProgram WindowsProgram WindowsProgram Windows ............................................................................................................................................................................................................................................................................................................................ 21212121

    3.4.1 .....Acquisition Control window (Single Mode) ....................................... 21 3.4.2 .....Acquisition Control window (Sequence Mode) ................................. 24 3.4.3 .....Camera Window ..............................................................................26 3.4.4 .....Acquisition Window .......................................................................26 3.4.5 .....Zoom Window ................................................................................ 27

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    4 Acquisition Program: Operation ....................................................................29 4.14.14.14.1 Logging inLogging inLogging inLogging in........................................................................................................................................................................................................................................................................................................................................................................ 29292929 4.24.24.24.2 Loading / Unloading the DrawerLoading / Unloading the DrawerLoading / Unloading the DrawerLoading / Unloading the Drawer ........................................................................................................................................................................................................................................ 30303030 4.34.34.34.3 Acquisition ConfigurationAcquisition ConfigurationAcquisition ConfigurationAcquisition Configuration ............................................................................................................................................................................................................................................................................ 30303030 4.44.44.44.4 Zoom ConfigurationZoom ConfigurationZoom ConfigurationZoom Configuration....................................................................................................................................................................................................................................................................................................................33333333 4.54.54.54.5 Acquisition Start / StopAcquisition Start / StopAcquisition Start / StopAcquisition Start / Stop ............................................................................................................................................................................................................................................................................................34343434 4.64.64.64.6 Gas Purge / PreparationGas Purge / PreparationGas Purge / PreparationGas Purge / Preparation............................................................................................................................................................................................................................................................................................35353535 4.74.74.74.7 During the AcquisitionDuring the AcquisitionDuring the AcquisitionDuring the Acquisition ................................................................................................................................................................................................................................................................................................ 36363636

    4.7.1......Details of the run ............................................................................. 36 4.7.2 .....Saving part of the acquisition .......................................................... 36 4.7.3 .....Windows Uses ................................................................................. 36

    4.84.84.84.8 Sequence ModeSequence ModeSequence ModeSequence Mode........................................................................................................................................................................................................................................................................................................................................37373737 4.8.1 .....Configuring and Starting a sequence :.............................................. 38 4.8.2 .....During a sequence : ......................................................................... 39 4.8.3 .....Stopping a sequence .......................................................................40

    5 Acquisition Program: Administration............................................................. 41 5.15.15.15.1 Account ManagerAccount ManagerAccount ManagerAccount Manager ................................................................................................................................................................................................................................................................................................................................ 41414141 5.25.25.25.2 Cassette ManagerCassette ManagerCassette ManagerCassette Manager ............................................................................................................................................................................................................................................................................................................................43434343 5.35.35.35.3 Export Log File /AuditExport Log File /AuditExport Log File /AuditExport Log File /Audit .................................................................................................................................................................................................................................................................................................... 44444444

    6 IMAGER: Maintenance ................................................................................. 45 6.16.16.16.1 TEA tank fillTEA tank fillTEA tank fillTEA tank fill ................................................................................................................................................................................................................................................................................................................................................................45454545 6.26.26.26.2 Argon cylinder replacementArgon cylinder replacementArgon cylinder replacementArgon cylinder replacement ................................................................................................................................................................................................................................................................ 48484848 6.36.36.36.3 Sample loader unblockingSample loader unblockingSample loader unblockingSample loader unblocking ........................................................................................................................................................................................................................................................................ 48484848 6.46.46.46.4 Remote help link (Telemaintenance)Remote help link (Telemaintenance)Remote help link (Telemaintenance)Remote help link (Telemaintenance)................................................................................................................................................................................................................ 49494949

    7 Material Safety Data Sheets (MSDS).............................................................. 50 TRIETHYLAMINE (TEA)TRIETHYLAMINE (TEA)TRIETHYLAMINE (TEA)TRIETHYLAMINE (TEA) ........................................................................................................................................................................................................................................................................................................................................................ 51515151 ARGONARGONARGONARGON ........................................................................................................................................................................................................................................................................................................................................................................................................................................54545454

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    Thank you for choosing the β IMAGER. This system is supplied complete : hardware and

    two software applications :

    β ACQUISITION for instrument control and image acquisition

    β VISION+ for image analysis and quantitation.

    Should you experiment difficulties, please contact Biospace lab Technical Department :

    Biospace lab

    10, rue Mercoeur 75011 PARIS – France

    tel : + 33.1.55.25.60.60

    fax : + 33.1.55.25.60.61

    e-mail : [email protected]

    This guide is a task-oriented user’s manual. It features both general and specialized information for advanced users.

    Important notice:Important notice:Important notice:Important notice:

    It is strongly advised to fully read this manual before launching your first acquisition. It is strongly advised to fully read this manual before launching your first acquisition. It is strongly advised to fully read this manual before launching your first acquisition. It is strongly advised to fully read this manual before launching your first acquisition.

    Afterwards, a content and index approach can be used to enquire about particular

    commands.

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    1 IMAGER introduction

    1.1 IMAGER description

    The β IMAGER originates from a technology initially developed for particle physics, and

    adapted to Beta labeling detection in biology by BIOSPACE and Nobel Prize winner Georges CHARPAK.

    It is primarily composed of a parallel-plate avalanche chamber (P.P.A.C.) -in essence the

    particle detector-, and of an acquisition system based on an intensified CCD camera with

    special optics. The chamber has an acquisition area of 200x250 mm². The zoom option allows an

    acquisition to be made of the whole area or of a particular region of interest of the sample

    (by automated camera repositioning), with improved spatial resolution.

    A gas mix composed of argon (from an external bottle) and Triethylamine (TEA for short; from a tank inside the equipment) is fed into the chamber. The proportions of this gas mix

    are computer-controlled; the computer also uses an array of sensors to monitor in real

    time the status of the equipment. This gas mix then has to be safely exhausted after it has

    been used.

    The acquisition computer on which the β ACQUISITION program is installed controls the

    entire system. This computer also hosts a modem used for remote assistance and diagnosis (Telemaintenance) by BIOSPACE. Data is usually analyzed on a networked

    computer using the β VISION VISION VISION VISION+ + + + program.

    The following two pictures highlight some β IMAGER elements and the usual on-site

    β IMAGER configuration.

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    ββββ IMAGER main elements

    Acquisition computer:

    Acquisition control & visualization

    Bubbler: gas flow visualization

    Control panel

    Sample loading

    assembly

    Location of the

    detector

    Acquiring system

    location

    (Intensified camera, optics,

    zoom)

    TEA tank location

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    Typical onTypical onTypical onTypical on----site site site site ββββ IMAGER IMAGER IMAGER IMAGER configuration configuration configuration configuration

    gas

    bottle

    gas exhaust

    BI

    OSPA

    CE

    2 ADDI-DATA cables

    and 2 PEGASUS cables

    regulator

    High grade Argon (4 bars)

    Control computer with

    ADDI-DATA and

    PEGASUS boards

    telemaintenance via modem

    phone network

    Ethernet network

    connection for empty bottle detection

    Analysis computer

    with Zip drive

    High resolution color printer (option)

    stabilized power supply (option)

    parallel cable

    power supply

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    1.2 IMAGER operating principles

    Samples are placed in a sample holder, which is automatically positioned inside the

    detection chamber. The detector is sensitive to all β emissions, including tritium, as well

    as all electron emitters such as 125I, 99mTc… all this with high efficiency.

    Any β particle entering the detector’s area ionizes the gas. An electrical field created by

    high voltages being applied onto the chamber grids accelerates primary ionization

    electrons thus freed. Successive electron collisions in the gas result in an avalanche of electrons and a scintillation light. The intensified CCD camera then records every light

    spot generated this way by every sample-emitted particle into the chamber.

    After the video signal from the camera has been digitized, a reconstruction algorithm

    allows the emission point to be calculated. Spatial co-ordinates from every particle are

    then stored this way. Counting of every detected particle gives, when accumulated, a real-

    time spatial distribution image of the sample activity.

    The β IMAGER allows both accurate spatial localization of radio labeled sites and direct

    quantification.

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    1.3 Control panel

    • POWERPOWERPOWERPOWER: This red light is on when the equipment is powered.

    • VOLTAGEVOLTAGEVOLTAGEVOLTAGE: This yellow light is on when an acquisition is started, and turns itself off

    when the detector is purged. At this point it is possible to open the sample loader to change samples, as soon as a sufficient overpressure is reached.

    • STAGE 1STAGE 1STAGE 1STAGE 1: Informs of sparking in the first detector stage.

    • STAGE 2STAGE 2STAGE 2STAGE 2: Informs of sparking in the second detector stage.

    • GENERAL RESETGENERAL RESETGENERAL RESETGENERAL RESET: This lights on when the sparks level safety limit is reached. When this occurs, the acquisition is automatically halted and a purge is launched. If this

    occurs, press the OK button on the message box that will appear and let the purge

    keep on. When it’s over, check the cleanliness of the sample.

    • DRAWERDRAWERDRAWERDRAWER, DRAWERDRAWERDRAWERDRAWER RESETRESETRESETRESET: This red light is on when the sample loader does not

    function properly (bad sample positioning, loader blocked). After correction, press the

    DRAWER RESET button in order to re-initialize the loader operation.

    • OPENOPENOPENOPEN DRAWERDRAWERDRAWERDRAWER: Button to open the loader.

    Important:Important:Important:Important: Do not open the loader until gas is circulating in the bubbler located on the

    front panel or until the status window indicates you can do it.

    NotNotNotNotice:ice:ice:ice: The β IMAGER latest generation is equipped with an electronic safeguard for this; for these machines, the above precaution is unnecessary, the loader being only enabled

    when the required conditions are met.

    CLOSECLOSECLOSECLOSE DRAWERDRAWERDRAWERDRAWER: Button to close the loader.

    • Check the sample holder positioning inside its groove in the support plate before closing the loader (sample holder translation should be

    impossible).

    • Also check there is nothing but the sample holder in the loader before

    closing it.

    • As a general precaution, do not leave the loader opened for too long to limit the introduction of dust into the detector.

    • If the equipment is not used for some time the loader can be closed without having a sample holder being loaded.

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    1.4 Operation precautions

    1.4.1 Avoid the following situations: Extreme temperatures (ideally between 20 and 25°C, tolerance 19° to 28°C)

    Direct sun exposition Rapidly changing temperature conditions

    Location on unstable or uneven surfaces

    1.4.2 Moving the equipment Never move the equipment while in oNever move the equipment while in oNever move the equipment while in oNever move the equipment while in operation or while a sample holder is loaded. peration or while a sample holder is loaded. peration or while a sample holder is loaded. peration or while a sample holder is loaded.

    Exit the software β ACQUISITION then make sure that both the β IMAGER and the

    computers are powered off before moving them. See the detailed procedure next page.

    To replace or move the Argon cylinder, always ensure the bottle is fully closed and

    that the pressure regulator is removed first.

    1.4.3 Gas exhaust Ensure that the exhaust tube is connected to an outside vent or a compatible Ensure that the exhaust tube is connected to an outside vent or a compatible Ensure that the exhaust tube is connected to an outside vent or a compatible Ensure that the exhaust tube is connected to an outside vent or a compatible exhaust extractor.exhaust extractor.exhaust extractor.exhaust extractor.

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    1.5 Normal operating conditions

    1.5.1 Power supply It is strongly advised to connect both the β IMAGER and the control computer to an uninterruptible power supply uninterruptible power supply uninterruptible power supply uninterruptible power supply (U.P.S.) to avoid power surges and cuts to interrupt an

    acquisition or even damage the system.

    The UPS must have a minimum power of 800VA. Its installation is described below.

    UPS installationUPS installationUPS installationUPS installation: The power plug must be connected to a well-earthed socket. The socket should only

    be used by the UPS; do not connect the UPS to extension leads or multiple sockets.

    Never disconnect the power lead while the UPS is on as this removes the earth from your equipment.

    Use the UPS to supply the β IMAGER and the control computer only. Please note that the average autonomy of the recommended UPS is 20 minutes.

    1.5.2 Turning on the β IMAGER • Open the gas cylinder

    • Switch on the β IMAGER

    • Switch on the control PC

    • Launch β ACQUISITION

    1.5.3 Turning off the β IMAGER • Do not switch of the β IMAGER while in acquisition.

    • Terminate the current acquisition.

    • Remove the sample holder from the machine and close the loader.

    • Exit the β ACQUISITION software.

    • Put the main power switch of the β Imager on the off position.

    • Close the gas cylinder.

    1.5.4 Characteristics of the β IMAGER • Electrical consumption: 170W max @ 220volts

    • Operating temperature: between +19 and +28°C.

    • Dimensions (HxLxW): 153cm x 63cm x 63cm

    • Weight: around 100 kg (+ 20 kg with the zoom option)

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    2 Sample handling

    2.1 Introduction

    The β IMAGER can handle several different types of sample holders designed for different types of samples:

    • Slides holding cassette: for thin anatomical slices on glass slides – standard cassette for 15 standard microscope slides (25x75 mm2 to 26x76 mm2).

    • Tritium sample holding cassette: for tritium-labeled samples: membrane, TLC plate, Whole Body Sections.

    • C14 sample holding cassette: for C14 -or any other medium to high-energy isotope- labeled samples on thin and flexible substrate: membrane, Whole Body

    Sections.

    2.2 Glass slides cassette

    This cassette is designed to hold 15 microscope slides with the following dimensions: 25x75 mm2 to 26x76 mm2, 1 mm thick. For other types of slides custom-built cassettes are

    used in the same way, only the number of slides held in the cassette might change.

    All sample holders come with a protective case. It must be kept clean (storage, periodical

    wipe with alcohol). A glass sample preparation plate of size 35x30 cm2 comes with this cassette. Thus the

    cassette can be manipulated on a flat & clean surface (periodically clean the plate with

    alcohol) before or after an acquisition.

    Handling principles:Handling principles:Handling principles:Handling principles:

    • Place the sample holder on the preparation plate (cavities underneath).

    • Unlock the lid and remove it.

    • Cover the backside of the slides with copper tape. Do not leave any copper tape on the edges of the slides or on the side of the sample.

    • Place all 15 slides in the cavities. All cavities need to be filled, by sample-free slides (with copper tape on one side) if necessary. The sample side must be

    against the preparation plate, the copper tape side being at the top.

    • Place the lid (springs underneath) on the sample holder, press the center of the lid and move all four knobs into the locked position.

    • Take the sample holder by the grooves in the locking knobs or by the holes in the lid depending on the version, and turn it around to check the absence of dust and to check the absence of dust and to check the absence of dust and to check the absence of dust and rough edges on the sample side.rough edges on the sample side.rough edges on the sample side.rough edges on the sample side.

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    NotesNotesNotesNotes: • It is recommended not to engrave or stick any labelnot to engrave or stick any labelnot to engrave or stick any labelnot to engrave or stick any label on the slides on the

    sample side. Or the back side of the sample-only the back side with no sample in front can be used to stick a label. FFFFlatness defaults alter the quality latness defaults alter the quality latness defaults alter the quality latness defaults alter the quality of the acquisitionof the acquisitionof the acquisitionof the acquisition.

    • The slides edges should be covered by the thin protective layer on top of the sample holder, no edges should be visibleno edges should be visibleno edges should be visibleno edges should be visible. (no broken slides)

    • The samples must be dry and held properly onto the slidebe dry and held properly onto the slidebe dry and held properly onto the slidebe dry and held properly onto the slide. • The use of some inksinksinksinks can also degrade the performance of the detector.

    • Open the drawer by pressing the "open drawer" button on the front panel of the machine and carefully place the sample holder in the groove or on the

    positioning knobs, according to the version.

    • The samples must be on the upper side.

    • Check that no translation of the sample holder is possible at this point.

    • Close the drawer and start an acquisition.

    2.3 Tritium (TLC) cassette

    There are two versions of this sample holder available:

    2.3.1 Basic version: Composed of a metal plate with a totally flat and smooth surface, with two bars

    underneath acting as feet.

    For tritium-labeled membranes or for tritiated tissues deposited on an adhesive isolating

    substrate – Do not use for TLCsDo not use for TLCsDo not use for TLCsDo not use for TLCs. Secure the sample with adhesive tape on the smooth surface so that it is as flat as

    possible.

    2.3.2 TLC version Composed of a metal support plate and of glass plates 2 to 4 mm thick fitting on the

    support plate.

    For tritium-labeled membranes & TLCs or for tritiated tissues deposited on an adhesive

    isolating substrate.

    • Thick TThick TThick TThick TLC plate LC plate LC plate LC plate (glass - thickness < 2,5 mm): simply fit the glass plate in its

    place (side to be analyzed on top) straight after a thorough drying of the sample.

    • Thin TLC plates Thin TLC plates Thin TLC plates Thin TLC plates (aluminum or flexible plastic - thickness < 0,5 mm): secure the plates on the supplied thin glass plate (dimensions 200x250 mm2 - thickness

    2 mm) with adhesive tape – place the glass plate + TLC assembly in the

    groove of the cassette (side to be analyzed on top) straight after a thorough

    drying of the sample.

    • MembranesMembranesMembranesMembranes (thickness < 0,5 mm): secure the membranes on a supplied thick glass plate (dimensions 200x250 mm2 – thickness 4 mm) with adhesive tape

    - place the glass plate + membrane assembly in the groove of the cassette

    (side to be analyzed on top)

    • Anatomical slices on isolating filAnatomical slices on isolating filAnatomical slices on isolating filAnatomical slices on isolating film m m m (thickness < 0,5 mm): secure the membranes on a supplied thick glass plate (dimensions 200x250 mm2 –

    thickness 4 mm) with adhesive tape - place the glass plate + slices assembly

    in the groove of the cassette (side to be analyzed on top)

    • Other types of saOther types of saOther types of saOther types of samples:mples:mples:mples: consult us

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    For both tritium cassettes, after the sample preparation steps described above:

    • Open the drawer by pressing the "open drawer" button on the front panel of the machine and carefully place the sample holder in the groove or on the

    positioning knobs, according to the version.

    • The samples must be on the upper side.

    • Check that no translation of the sample holder is possible at this point.

    • Close the drawer and start an acquisition in in in in modemodemodemode corresponding to Tritium corresponding to Tritium corresponding to Tritium corresponding to Tritium

    cassetcassetcassetcassette + Low resolutionte + Low resolutionte + Low resolutionte + Low resolution (the High reso (the High reso (the High reso (the High resolution mode is not allowed this lution mode is not allowed this lution mode is not allowed this lution mode is not allowed this cassette)cassette)cassette)cassette)

    Handling precautions:Handling precautions:Handling precautions:Handling precautions:

    • Always check that the samples are well secured onto the glass plate, nothing nothing nothing nothing should be sticking out more than 1mmshould be sticking out more than 1mmshould be sticking out more than 1mmshould be sticking out more than 1mm above the cassette’s level to avoid any contacts with the detector’s grid (be especially careful with rat slices)– If the sample was to stick out, the use of a thinner glass plate generally allows an

    acquisition to be made (background noise might be slightly higher).

    • The samples must always be pppperfectly driederfectly driederfectly driederfectly dried, as traces of humidity or solvents can alter the operation of the machine and could even degrade it. TLC plates must . TLC plates must . TLC plates must . TLC plates must always be dried just before analysisalways be dried just before analysisalways be dried just before analysisalways be dried just before analysis (with a heating plate or a hot air gun), since they systematically pick up humidity.

    • While removing the samples from the plate, be careful not to scratch the platenot to scratch the platenot to scratch the platenot to scratch the plate. Clean the plate regularlyClean the plate regularlyClean the plate regularlyClean the plate regularly to remove any adhesive tape or glue.

    • When you change samples, it is recommended not to leave the loading drawer not to leave the loading drawer not to leave the loading drawer not to leave the loading drawer opened for too longopened for too longopened for too longopened for too long to avoid any risk of dust depositing on it and thus getting into the detector.

    • This cassette is designed for tritiumtritiumtritiumtritium, which is a very low energy isotope. . . . The use of this cassette with higher energy isotopes (14C, 35S, 33P...) does not allow as

    good a spatial resolution as for tritium. It is recommended for these isotopes to use the dedicated cassettes available.

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    2.4 C14 (Non-tritiated membrane) cassette

    Sample holder exclusively designed for very thin samples on thin & flexible substrateexclusively designed for very thin samples on thin & flexible substrateexclusively designed for very thin samples on thin & flexible substrateexclusively designed for very thin samples on thin & flexible substrate, such as membranes or tissue microtomes on flexible plastic film (Whole Body Sections)

    labeled with a medium or high-energy isotope (14C, 35S, 33P, 32P, …). This sample holder is composed of a hollow metal frame on which a very thin grid and a

    metallized Mylar sheet are tensed, with a pressing plate that fits in the previous frame to

    press the samples against the metallized Mylar. It is supplied with a plastic protective

    plate which fits on the grid side of the cassette, a storage case, a roll of adhesive film to

    protect and clean the grid and finally a very thin (3 µm) Mylar roll to wrap the samples with

    to protect the cassette’s Mylar sheet from any contamination or marking.

    This extremely delicate sample holder must be handled with care and must be stored in This extremely delicate sample holder must be handled with care and must be stored in This extremely delicate sample holder must be handled with care and must be stored in This extremely delicate sample holder must be handled with care and must be stored in its case after use. its case after use. its case after use. its case after use.

    Handling principles:Handling principles:Handling principles:Handling principles:

    • Remove the cassette + protective plate assembly from the storage case.

    • Place this assembly with the cassette upside on a table.

    • Open the cassette by lifting the pressing plate once all four locking pins are

    unlocked.

    • A thin protective Mylar sheet should already be inside to protect the cassette’s

    own Mylar window – if it is clean you may use it, otherwise replace it by a new sheet leaving some extra material on the sides – the whole window surface

    must be covered by the protective Mylar sheet.

    • Place the samples on this protective Mylar sheet, so that they are flat, that they are not superimposed and that the side to be analyzed is looking downwards

    (against the protective Mylar sheet).

    • Fold back the Mylar onto the samples.

    • Place the pressing plate in its position (smooth side underneath) and press

    lightly in its center to move the locking pins into the right position.

    • Holding the cassette by its sides, turn it around.

    • Place the cassette into the groove in the drawer’s support plate, grid upward.

    • Check that no translation is made possible.

    • Remove the protective film from the grid to place it onto the cassette’s

    protective plate still on the table to protect it during the acquisition.

    • Close the drawer and start an acquisition in the "Cassette C14" High Resolution mode.

    Note:Note:Note:Note: The "Low Resolution" mode can also be used with this cassette for counting purposes, but the resolution degradation is important.

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    Once the acquisition is finished:

    • Open the drawer

    • Carefully stick a new protective film onto the cassette grid.

    • Remove the cassette, and turn it around to place it onto the protective plate.

    • Remove the samples and their Mylar sheet.

    • Place a new Mylar sheet into the cassette.

    • Place a new sample in or close the cassette and place the cassette + protective plate in the storage case. (grid and Mylar side always against protective glass

    plate )

    Handling precauHandling precauHandling precauHandling precautions:tions:tions:tions:

    � Never lay the C14 cassette elsewhere than on the cassette protective platecassette protective platecassette protective platecassette protective plate or on the β IMAGER drawer support platedrawer support platedrawer support platedrawer support plate.

    � A protective film must always cover the grid of the C14 cassette A protective film must always cover the grid of the C14 cassette A protective film must always cover the grid of the C14 cassette A protective film must always cover the grid of the C14 cassette until the drawer of the β IMAGER is to be closed to start an acquisition.

    � Always remove the protective film before closing the drawer of the Always remove the protective film before closing the drawer of the Always remove the protective film before closing the drawer of the Always remove the protective film before closing the drawer of the β IMAGER. . . . � A protective Mylar sheet must always be present inside the C14 cassette (even when

    stored and without a sample in)

    � The cassette’s Mylar window and grid are delicate: samples should not show any samples should not show any samples should not show any samples should not show any sharp edgessharp edgessharp edgessharp edges that might damage them - ScrupulouslScrupulouslScrupulouslScrupulously follow the above handling y follow the above handling y follow the above handling y follow the above handling procedureprocedureprocedureprocedure.

    � The Mylar window can be carefully cleaned carefully cleaned carefully cleaned carefully cleaned with a lint-free paper soaked in alcohol, although parsimoniously to avoid removing the metallization.

    � Should a hot spothot spothot spothot spot be present during the acquisition, abort the acquisition, open the drawer and place a new adhesive film on the grid to clean it, remove it, and then

    restart. If the problem is persistent check the sample.

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    2.5 Sample – Cassette matching

    Use the correctcorrectcorrectcorrect sample holder for the sample to be acquired:

    Isotopic label Isotopic label Isotopic label Isotopic label

    SampleSampleSampleSample 3H3H3H3H

    High energy isotopeHigh energy isotopeHigh energy isotopeHigh energy isotope

    (14C, 33P, 35S, 32P...)(14C, 33P, 35S, 32P...)(14C, 33P, 35S, 32P...)(14C, 33P, 35S, 32P...)

    MembranesMembranesMembranesMembranes 3H cassette + 4mm glass plate

    14C cassette + Mylar protection 3µm

    Thin TLC *Thin TLC *Thin TLC *Thin TLC *

    Thickness < 500µm

    3H cassette + 2mm glass plate

    3H cassette + 2mm glass plate

    Note: degraded resolution

    Thick TLC *Thick TLC *Thick TLC *Thick TLC *

    Thickness 1 to 2 mm

    3H cassette

    3H cassette

    Note: degraded resolution

    Whole Body SectionWhole Body SectionWhole Body SectionWhole Body Section 3H cassette + 2/4mm glass plate

    14C cassette + Mylar protection 3µm

    Glass slidesGlass slidesGlass slidesGlass slides

    Slide cassette

    High Resolution Mode or not,

    according to sample quality

    Slide cassette

    High Resolution Mode or not,

    according to sample quality

    OthersOthersOthersOthers Ask for adviceAsk for adviceAsk for adviceAsk for advice

    *: For TLC, thorough drying of the sample is necessary just before acquisition.*: For TLC, thorough drying of the sample is necessary just before acquisition.*: For TLC, thorough drying of the sample is necessary just before acquisition.*: For TLC, thorough drying of the sample is necessary just before acquisition.

    Important general remarks:Important general remarks:Important general remarks:Important general remarks:

    • Analyze only dry, cleandry, cleandry, cleandry, clean and wewewewellllllll----fixedfixedfixedfixed samples (no particles or dust)

    • Sample emerging activity must not exceed 2kBqmust not exceed 2kBqmust not exceed 2kBqmust not exceed 2kBq, in full field.

    • Make sure that the sample is flatflatflatflat

    • For the regulation to work properly, the samples in the acquisition area should have

    a minimum β IMAGER detected activity around 100 cpm

    • For quantificationquantificationquantificationquantification purposes, it is recommended to add one standard of the same to add one standard of the same to add one standard of the same to add one standard of the same isotopeisotopeisotopeisotope as the samples’ and of a known activity, in order to allow the conversion of β IMAGER -detected cpm /mm² into the more common dpm/mg format.

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    3 Acquisition Program: Presentation

    3.1 Introduction

    This new software is designed to work with Windows 2000.

    Unlike its predecessor, it is fully integrated in Windows and benefits from it.

    Familiarity with this program is not enough to run the β IMAGER; it is essential to be familiar also with sample handling for this instrument, since it is a critical part to get the

    best images from the β IMAGER.

    The β ACQUISITION software can operate in two different modes:

    • Single Mode : you configure and run one acquisition after the other

    • Sequence Mode: you configure a sequence of acquisitions and start the sequence that will automatically run the programmed acquisitions one after the

    other. This can be very useful for zooming on different slides after a full field

    acquisition for example.

    The next sections explain how to use the β IMAGER with these two operating modes.

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    3.2 Menus

    3.2.1 File Menu This menu is shown below:

    This menu allows the user to:

    • Open a previously acquired acquisition in the Zoom Window

    • Save a currently running (or last acquired) acquisition for immediate analysis

    • Log Out thus change the User identification for the next acquisition or for any further event appearing in the log file

    • Lock Session (C.f. Account Manager section) to stop a different User from logging

    in after a time out

    • Exit β−ACQUISITION

    3.2.2 Tools Menu This menu is shown below:

    This menu allows the user to:

    • Change the password for the active login account

    This menu allows the administrator to:

    • Manage the correction files for each sample holder

    • Manage/Create the logging accounts & their access levels for β−ACQUISITION

    • Export the log files, tracing events, errors & acquisitions for the instrument

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    3.2.3 View Menu This menu is shown below:

    The Overview Panel provides information on the current state of the instrument for service purposes and, when an acquisition is running, the “Quality” criterion provides information on how well the acquisition is running.

    The “Quality” criterion is getting close to zero when the acquisition regulation is stable, i.e. the lower this criterion, the better the acquisition.

    We recommend discarding acquisitions with a “Quality” criterion higher than 0.5 as the acquisitions would not have been stable enough.

    This criterion is available in the properties of the image when the acquisition is opened

    using the analysis program.

    3.2.4 Help Menu This menu allows the user to display the version of β−ACQUISITION is currently installed by clicking on About.

    3.3 Shortcut Icons

    The shortcut toolbar is shown below:

    Allows to save the current acquisition "as is" while the acquisition keeps running

    Displays / hides the camera window

    Displays / hides the acquisition window

    Displays / hides the Zoom window

    Hides every image window

    Reorganizes the program windows

    Information about the software

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    3.4 Program Windows

    3.4.1 Acquisition Control window (Single Mode)

    TEA level indicator Information about

    running acquisition

    Start / Stop acquisition

    Sample Holder

    selection

    Isotope Selection

    Zoom Selection

    Stop Conditions

    (optional)

    Acquisition Name

    & Path

    Comments (optional)

    TEA tank fill

    Acquisition mode

    selection

    Update the Zoom

    Configuration

    Load background image

    in Zoom Window

    Current status LED Current status

    information

    Sequence Mode /

    Single Mode

    Gas Preparation / Purge

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    Current status LED: (information)

    Green when the instrument is in standby & TEA has been purged

    Red in acquisition

    Yellow when TEA has not been purged

    Current status information text box:

    Provides information about the phase the β IMAGER is in. Error conditions are also displayed here. (time left to the end of an action)

    Acquisition time / Total count / Counts per minute : (information) Self-explanatory – The counts or time left to the stop condition can be obtained by

    clicking on the item.

    TEA level indicator : Displays the current level of TEA in the instrument.

    Green when the level is sufficient

    Orange when a TEA tank fill can be done

    Red when the level is too low to start an acquisition

    TEA Fill button : (Advanced User access level required)

    Grayed when a TEA tank fill is not necessary (green TEA level indicator) or not

    allowed (during acquisition or in sequence mode for example), otherwise this button is enabled.

    For a TEA tank fill please refer to the "TEA Tank fillTEA Tank fillTEA Tank fillTEA Tank fill" section in the "MaintenanceMaintenanceMaintenanceMaintenance" chapter.

    Start New / Stop button :

    To start an acquisition once it has been configured, or to stop a running

    acquisition.

    Preparation / Purge button :

    Preparation button : Used to prepare the detector to an acquisition; the acquisition

    gas mix will be flown through the detector.

    Purge button : To flush out the TEA from the detector in order to remove any sample holder.

    Isotope selection : (Compulsory, configuration)

    Select the correct isotope from the list to configure an acquisition with the right parameters.

    (If you use an isotope not listed please contact Biospace)

    Sample holder selection : (Compulsory, configuration) Match the sample holder you loaded into the machine: DO use the right sample

    holder, this is critical.

    Acquisition mode selection : (Compulsory, configuration) Available acquisition modes with the selected sample holder.

    Zoom Mode : (Compulsory, configuration)

    � Zoom position (unrolling list): choose one of four preset zoom position, & adjust

    the position of the zoomed area in the "Zoom Window".

    � Load file button: allows you to select a background image (a previous full field

    acquisition) for a more precise zoom positioning. � Change configuration button: no changes on the zoom are taken into account

    until you press this button (just before starting an acquisition)

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    Stop conditions : (Optional, configuration)

    Acquisitions can be stopped automatically after a set time or a set count.

    "Use stop conditions" has to be checked and "Apply" clicked before those stop conditions are taken into account.

    Purge on stop allows if unchecked to chain acquisitions of the same sample

    without having to purge & prepare the gas mix between acquisitions.

    Acquisition name & path :

    A name including the date and an index is proposed as a default name but can be

    changed. The *.BVR image file and the associated *.CRI and *.XLS are recorded in a

    common folder with the same name.

    Comments :

    Self-explanatory

    These comments will be part of the *.BVR image file.

    Sequence Mode / Single Mode button :

    Press Sequence Mode (when in Single Mode) to use a sequence of acquisition

    performed automatically one after the other. (See Sequence Mode section for more details)

    Press Single Mode (when in Sequence Mode) to perform acquisitions manually one

    after the other.

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    3.4.2 Acquisition Control window (Sequence Mode) When you go to Sequence Mode, some new controls appears into the Acquisition Control

    window :

    Total number of

    acquisition in sequence

    Acquisition Number of the acquisition being

    configured number

    Sequence Mode /

    Single Mode

    Add the configured

    acquisition to sequence

    Run the Next

    acquisition in

    sequence

    Reset sequence

    Current running acquisition number

    Configuration of the acquisition as selected

    in Acquisition Number

    Load / Save sequence

    configuration

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    Current Acquisition number :

    Position in the sequence of the running acquisition .

    Total Acquisitions : Number of acquisitions configured in the sequence

    Acquisition Number :

    Position of the current acquisition being configured within the sequence. Once an acquisition configuration has been added to a sequence , it appears between

    brackets in the list.

    “Add to Sequence” button : To add the configured acquisition to the sequence (or to record any change in this

    configuration)

    “Load / Save Sequence” button : To use a previously saved configuration (sequence): once the sequence is saved it

    can be recalled as often as required.

    “Reset Sequence” button : To reset the sequence that is to say to remove all the configured acquisition from

    the sequence

    “Single Mode” button : To go back to single acquisition mode

    “Run Next” button : To run the next acquisition in the sequence when you stop manually the previous acquisition, thus avoiding resuming the acquisition & re-enter stop conditions to

    continue the sequence.

    Important: A change to an acquisition configuration is only taken into account in the sequence if the “Add to Sequence” button is clicked after making the change.

    For other controls of the window see section Acquisition Control window (Single Mode).

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    3.4.3 Camera Window This window allows visualizing the events the way the camera detects them: as

    disintegrations.

    This window may be resized by dragging its edge while holding the mouse left button down.

    It can be used:

    � To check the activity or activity distribution of the sample � To detect non-radioactive hotspots: almost always present emitting points that

    bias the amplification regulation (because they are not coming from

    radioactivity). These usually come from fibers, hair, or large dust specks on the

    sample. They also often create sparks (violent discharges in the detector).

    Right click in the window area to modify the display palette (color scheme).

    There are five available palettes:

    � Gray: linear palette, from black to white � Invertgray: linear palette, from white to black

    � BlackWhite: binary palette, either black (nothing) or white

    � Gamma: logarithmic palette, from black to white (brings out low levels thus used

    for low levels) � Rainbow: linear palette, from black to red (colored)

    � Camera: linear palette, from black to red (colored) – same as before but

    simplified

    To detect non-radioactive hotspots, the best palettes are BlackWhite and Camera.

    Note: The Camera window display is very demanding in computer resources. For this reason, please avoid enlarging the image too much & leaving it so.

    3.4.4 Acquisition Window This window displays the accumulation of every acquired event since the beginning of the acquisition. In effect, this can be seen as what the autoradiographic image will look like.

    This window may be resized by dragging its edge while holding the mouse left button

    down.

    Right click in the window area to modify the display palette (color scheme).

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    There are five available palettes:

    � Gray: linear palette, from black to white

    � Invertgray: linear palette, from white to black

    � BlackWhite: binary palette, either black (nothing) or white � Gamma: logarithmic palette, from black to white (brings out low levels thus used

    for low levels)

    � Rainbow: linear palette, from black to red (colored)

    � Camera: linear palette, from black to red (colored) – same as before but simplified

    Right click in the window area to modify the display scale (scale range).

    It can be left as automatic (checkbox checked) or the maximum & minimum be manually

    adjusted by either using the sliding bars or by replacing the values in the corresponding boxes and pressing the enter key.

    Note: The window display is very much simplified since such an item is very demanding in

    resources. For this reason the image will have a much better resolution in the analysis

    software but can also have a "cloud of points" effect if there are not enough statistics

    3.4.5 Zoom Window If your β IMAGER has the zoom option, you have the possibility to physically zoom in a

    particular region of the acquisition area.

    Four zoom positions are available, and can be selected from the Control Window:

    � Full field (200x250 mm2 or 15 slides)

    � 6 Slides (120x160 mm2)

    � 2 Slides (75x100 mm2) � 1/2 Slides (25x32 mm2)

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    When the zoom is not in full field, a rectangle representing the area to acquire appears in the Zoom Window. This rectangle can be positioned precisely by holding the left mouse button while over the rectangle & moving the mouse to the required position.

    This window may be resized by dragging its edge while holding the mouse left button

    down.

    It is also possible to change the background image of this window by pressing "Load" in the Control Window and selecting the full field image to be used in the appearing dialog

    box.

    Right click in the window area to modify the display palette (color scheme).

    There are five available palettes:

    � Gray: linear palette, from black to white � Invertgray: linear palette, from white to black

    � BlackWhite: binary palette, either black (nothing) or white

    � Gamma: logarithmic palette, from black to white (brings out low levels thus used

    for low levels)

    � Rainbow: linear palette, from black to red (colored) � Camera: linear palette, from black to red (colored) – same as before but

    simplified

    Right click in the window area to modify the display scale (scale range). It can be left as automatic (checkbox checked) or the maximum & minimum be manually

    adjusted by either using the sliding bars or by replacing the values in the corresponding

    boxes and pressing the enter key.

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    Note: No changes are applied to the zoom configuration until you press "Change Change Change Change ConfigurationConfigurationConfigurationConfiguration" in the Control Window. This can be done just before starting the acquisition Note: DO wait that the zoom has finished moving before starting an acquisition.

    4 Acquisition Program: Operation

    4.1 Logging in

    When the acquisition program is started, the user will be asked to identify before being

    allowed to go any further.

    Every user has to be registered by the administrator of the instrument using the Account Manager tool, accessible via the Tool menu by the administrator. When a different user starts using the instrument while the acquisition program is

    running, the current user can log off via the File / log off menu.

    The logging window is shown below:

    When the Cancel button is clicked, the acquisition program terminates. If the user is logging in for the first time, or if the password expired (c.f. the Administration

    section), the user is asked to enter a different password.

    Note: The Login & Password are case-sensitive.

    There are 3 access levels available:

    • UserUserUserUser: to start, stop configure an acquisition & a sequence.

    • Advanced UserAdvanced UserAdvanced UserAdvanced User: to refill the TEA and do all of the above.

    • AAAAdministratordministratordministratordministrator: To manage users, correction files export log files and do all of the above.

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    4.2 Loading / Unloading the Drawer

    When the acquisition program is started or when an acquisition is stopped, a detector

    purge is launched if it was not done or completed before.

    Also, the zoom is indexed every time the program is launched.

    If the detector is purged, its pressurization is initiated, and the status box in the Control

    Window displays "Pressurizing detector".

    Once the detector has reached the required pressure, the instrument goes into a waiting

    state (Ready for opening or starting an acquisition in the status box).

    At this point the drawer can be opened by pressing the Open loader button at the front of the instrument.

    It can then be loaded or unloaded as required.

    Note :If the instrument has not been used for a long time, hold the drawer closed with your hand until it goes into alarm (red LED at the front of the instrument) to allow the loader support plate to fall in place: the seal of the detector might retain the plate longer than expected if it was not exercised for a period of time! If this happens the drawer may be blocked and the seal may be ripped! The drawer alarm can be cleared by pressing the Clear Drawer Alarm button at the front of the instrument. Once loaded, the drawer can be closed by pressing the Close loader button at the front of

    the instrument.

    Note: Do not hesitate to hold the drawer if the cassette is not positioned properly in its groove! The detector may be damaged and the drawer may be blocked if you fail to do so! The drawer alarm can be cleared by pressing the Clear Drawer Alarm button at the front of the instrument.

    4.3 Acquisition Configuration

    Before an acquisition is started, the correct acquisition parameters have to be entered by

    the user. They are critical for a good acquisition since they are not just information, but

    are used for the regulation of the acquisition.

    In the Control Window:

    � Choose the isotope (Isotope box)

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    � Choose the cassette or sample holder (in Sample Holder, click on the required sample holder);

    The available sample holders then appears in a menu: choose the correct one.

    Note: The selected sample holder name will then be highlighted by a check symbol.

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    � If required, change the Acquisition Mode, which is as a default the optimal setting for the selected cassette : Low Resolution for 3H cassette (the only mode

    compatible with this cassette) and High Resolution for the other cassettes.

    Change this setting to Low Resolution if you experience sparks : this low-resolution mode is more tolerant to not-so-perfect samples (flatness, cleanliness)

    but as its name indicates the resolution is poorer, especially with other isotopes

    than Tritium.

    � As an option, stop conditions can be entered either as a set time in hours &

    minutes or as a required count. Once this is done, " Use stop conditions " must be checked & the " Apply " button must be pressed to take these stop conditions

    into account.

    Notes : Stop conditions can be changed while the acquisition is running, by entering the new values followed by " Apply ". You can use both stop conditions in time and count at the same time, the first condition to be reached will stop the acquisition.

    • If another acquisition will be run on the same sample (without removing it from the instrument), the detector purge & gas preparation can be skipped by

    unchecking " Purge on Stop ".

    • The name & path of the acquisition file to be acquired are automatically

    generated. However, they can be changed in the " Acquisition Name " box.

    • Comments can also be entered & will be recorded with the file. (Up to 256 characters).

    At this point, the zoom can be configured or the acquisition started.

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    4.4 Zoom Configuration

    Before zooming in a part of the acquisition area, it is essential to run a full field

    acquisition to ensure of the absence of non-radioactive hotspots (they can be detected

    with the Camera Window), which are induced by the presence of fibers, hair, flaking samples, large dust specks.

    Those non-radioactive hotspots -besides being non-desirable signals- can alter the

    regulation of the acquisition if too important proportionally to the radioactive signal (not being radioactive, they have different characteristics from the expected signal with which

    the regulation is made), and they can create sparks (violent electrical discharges in the

    detector).

    Those non-radioactive hotspots can be detected since they are almost always present in

    the Camera Window, unlike radioactive signal.

    The zoom configuration parameters can be changed in the " Zoom Mode " part of the

    Control Window in conjunction with the Zoom Window ( icon).

    Four zoom positions are available, and can be selected from the Control Window:

    � Full field (200x250 mm2 or 15 slides)

    � 6 Slides (120x160 mm2) � 2 Slides (75x100 mm2)

    � 1/2 Slides (25x32 mm2)

    Note: The default Zoom mode when the acquisition program is started is Full Field. Therefore an acquisition can be started without configuring the zoom if the zoom options did not change.

    To ease the zoom positioning, a full field image of the sample to be acquired can be

    loaded as a background image for the Zoom Window, by pressing " load file " in the Control Window.

    When this done, the following dialog window appears:

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    The required BVR file to be used as a background image can then be selected to appear in

    the Zoom Window.

    Right clicking anywhere in the Zoom Window allows you then to change the color palette

    or the scale.

    When the zoom is not in full field, a rectangle representing the area to acquire appears; This rectangle can be positioned precisely by holding the left mouse button down while over the rectangle & moving the mouse to the required position.

    In Single Mode : In Single Mode : In Single Mode : In Single Mode :

    Once the zoom is fully configured press "Change ConfigurationChange ConfigurationChange ConfigurationChange Configuration" button to validate it. The

    zoom is then automatically positioned and you can start the acquisition (if you have

    configured the other parameters for the acquisition)

    In Sequence Mode : (see Sequence MoIn Sequence Mode : (see Sequence MoIn Sequence Mode : (see Sequence MoIn Sequence Mode : (see Sequence Mode section)de section)de section)de section)

    The zoom is automatically positioned for each acquisition of the sequence – you don’t need to press "Change ConfigurationChange ConfigurationChange ConfigurationChange Configuration" button.

    4.5 Acquisition Start / Stop

    Before starting an acquisition, it has to be configured.

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    Once the acquisition has been configured (including zoom configuration and its validation

    with "Change Configuration" button), pressing "Start new" button in the Control Window will start the acquisition.

    As soon as an acquisition is in progress (including the gas preparation phase), the text of

    the "Start new" button toggles to " STOP ". The acquisition can thus be stopped at any time (even before any event was acquired i.e.

    during gas preparation). The acquisition can also stop automatically on Stop Conditions if selected and applied.

    4.6 Gas Purge / Preparation

    According to the phase the β IMAGER is in, the "Purge / Preparation " button in the Control Window will be available as "Purge" or as "Preparation".

    If the gas mix in the detector contains TEA (gas preparation or ready for an acquisition), the available button is "Purge" and by pressing it the TEA will be cleared from the detector to allow the drawer to be opened.

    If the gas in the detector is only argon (or argon + air after drawer opening), the available button is "Preparation" and by pressing it the detector gas mix will be prepared for acquisition; the acquisition parameters can be entered following this thus saving a few

    minutes.

    Once the gas preparation is launched, pressing "Start New" will start the acquisition as soon as the preparation is completed.

    At any rate pressing "Start new" will launch a preparation for your first acquisition.

    If the acquisition program was closed without having completed a detector purge, it will automatically do so when the program is restarted.

    To avoid purging when the sample is not to be removed, the "Preparation" button can be used immediately.

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    4.7 During the Acquisition

    4.7.1 Details of the run During the acquisition you can read on the Control Panel : � Acquisition time (or Time Left by clicking on the value if stop conditions are used).

    � Total count (or Remaining Count by clicking on the value if stop conditions are used).

    � Count per minute ( 100 < cpm < 120000)

    4.7.2 Saving part of the acquisition The data already acquired during an acquisition can be exported & analyzed as a new

    acquisition file without having to interrupt the run, simply by clicking on the icon. A dialog window then appears indicating the progression of the save process.

    The file created in this manner can immediately be moved & analyzed.

    Once the acquisition has been stopped (even after a program restart) this functionality is

    still available.

    4.7.3 Windows Uses

    The Camera Window ( ) allows the user to:

    • Check the presence of non radioactive hotspots (points permanently present)

    • See the activity distribution of the sample (s)

    The Acquisition Window ( ) allows the user to see the events already acquired.

    The Acquisition Control allows the user to see time and count information on the

    acquisition, to check the instrument status, to modify the stop conditions and to interrupt

    the acquisition.

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    4.8 Sequence Mode

    The β ACQUISITION software allows an acquisition to be pre-programmed and run as a

    sequence (up to fifty consecutives acquisitions) to be configured and customized by the user. This can be of particularly useful to automatically acquire zoom images after a full

    field acquisition.

    The following sections describe the different functions that are specific to the sequence mode. Details regarding the main menu and the default dialog box (single mode) are still

    relevant to sequence mode version. The Sequence mode is controlled in the same control

    window as the usual Single Mode.

    To use the Sequence Mode, first press the "Sequence Mode" button.

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    4.8.1 Configuring and Starting a sequence :

    To configure the acquisitions in the sequence :

    1. Select the Acquisition Number

    Note: the sequence will run the acquisitions configured and added to sequence from number 1 to the last number configured in increasing number order

    2. Configure this acquisition as in Single Mode

    • Select the isotope

    • Select the sample holder

    • Select the acquisition mode & correction

    • Insert the stop conditions for this acquisition

    • Select the Zoom mode and position for this acquisition

    • Give a name and location (optional)

    • Add comments (optional)

    Note: Stop conditions are mandatory in the sequence acquisition mode

    To have a configured acquisition included in the sequence :

    3. Press the "Add To Sequence" button

    To configure and add to sequence the next acquisitions you want to run the same way :

    4. Select a new acquisition number (position) 5. Configure the acquisition

    6. Add the acquisition to the sequence

    Once all acquisitions have been configured and added in the sequence, the programmed runs can be launched by:

    7. Press the "Start New" button.

    The sequence will then begin with acquisition n° 1. When stop conditions are

    reached it will automatically run the next acquisition in the sequence and so on.

    Note : You may modify all of your settings while programming the sequence, by selecting the acquisition to modify, change it and press the "Add to sequence" button again.

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    Important : Important : Important : Important :

    Before running a sequence with zoom in a part of the acquisition area, it is

    essential to run a full field acquisition to ensure of the absence of non-

    radioactive hot spots (they can be detected with the Camera Window), which

    are induced by the presence of fibers, hair, flaking samples, large dust specks. Those non-radioactive hotspots -besides being non-desirable signals- can alter the

    regulation of the acquisition if too important proportionally to the radioactive signal (not

    being radioactive, they have different characteristics from the expected signal with which

    the regulation is made), and they can create sparks (violent electrical discharges in the

    detector).

    Those non-radioactive hotspots can be detected since they are almost always present in the Camera Window, unlike radioactive signal.

    4.8.2 During a sequence : The number of the running acquisition can be seen in the "Current acquisition" box. As in

    Single Mode the configuration and the time and counting information of the running

    acquisition are set in the Acquisition Control window. The zoom selected area is set in the

    Zoom window. The image is displayed in the Image window.

    The “Stop” button can be pressed without having to wait for the stop conditions to be fulfilled. (for example, if you want to modify the acquisitions parameters).

    From an interrupted sequence it is possible to run the next acquisition of the sequence by

    pressing the "Run Next" button.

    To modify acquisition settings : stop the running acquisition, select the number of the acquisition to be modified, modify it and press the "Add to sequence" button.

    During the sequence you can modify the stop conditions of the running acquisition by

    changing them and pressing the "Apply" button in the Stop Conditions zone as you would in Single Mode. To change the stop conditions of another acquisition in the sequence you have to stop the sequence, change the stop condition and restart the sequence as

    explained before.

    In the sequence mode, you cannot use the “Purge / Preparation” button, and you cannot launch the TEA Fill procedure. In order to do this, you will have to stop the sequence and

    go back in the single acquisition mode, by pressing the "Single Mode" button. Once in single acquisition mode, you will be able to go back in the sequence mode.

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    4.8.3 Stopping a sequence

    When the last acquisition in the sequence is over the sequence is automatically

    stopped.

    If you check the purge checkbox in the stop conditions of the last acquisition, a

    purge automatically follows the acquisition. When the purge is over you can

    then remove your sample as in Single acquisition Mode.

    If you didn't set a purge in the stop conditions of the last acquisition the β

    IMAGER is waiting for another acquisition. You can then :

    � Reset the sequence, program a new sequence and run it on the same samples

    � You can also go back to Single Mode and start a new acquisition

    When no automatic purge is programmed going back to Single Mode to purge

    manually allows the removal of the samples.

    Note : You can stop any acquisition of the sequence at any time with the Stop

    button

    Note : You cannot launch twice the same sequence.

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    5 Acquisition Program: Administration

    5.1 Account Manager

    Every user has to be registered by the administrator of the instrument using the Account Manager tool, accessible via the Tool menu by the administrator before being allowed to operate the instrument.

    The Account Manager tool window is shown below:

    There are 3 access levels available:

    • UserUserUserUser: to start, stop configure an acquisition & a sequence.

    • Advanced UserAdvanced UserAdvanced UserAdvanced User: (or Calibration User) to refill the TEA and do all of the above.

    • AdministratorAdministratorAdministratorAdministrator: To manage users, correction files export log files and do all of the above.

    The Account Manager tool allows the administrator to create, revoke logins as well as set the duration for the validity of the password used for the accounts & their inactivity time

    out. (The user can change his/her password at all times nonetheless)

    Checking “Lock current session” locks the possible logins on the current login during time out (inactivity automatic lock or log out set in the same window).

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    When the administrator creates a new login account the following window appears:

    A temporary password has to be assigned : the new user will be prompted to change the

    password on first use before being allowed to go any further.

    A comment is associated with this account creation.

    When the administrator revokes a login account, a comment is associated with the

    deletion (“Reason for revocation”: window appears on clicking the Revoke button).

    Notes: When a different user starts using the instrument while the acquisition program is running, the current user can log off via the File / log off menu. If the user is logging in for the first time, or if the password expired, the user is asked to enter a different password before being allowed to go any further. The Login & Password are case-sensitive. The Password has to be alpha-numerical & of at least 8 characters.

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    5.2 Cassette Manager

    In order to reduce unhomogeneity of the acquisition field of view (compulsory in some

    acquisition modes), a correction is applied in real time to the acquisition data; this

    correction is reversible & is specific for a sample holder (cassette) in its designed operating mode.

    Generating & managing the homogeneity correction files is usually done by Biospace lab

    or its Representative: what follows is informational & does not require maintenance by the instrument administrator.

    There are 3 operating modes corresponding to a different type of sample holder:

    • Tritium cassette

    • Slides cassette

    • High Energy membrane cassette (14C cassette)

    A correction file has to be associated with each cassette (i.e. if you possess several

    cassettes of the same type, you need to use as many different correction file); this is done

    via the tool shown below, the Cassette Manager tool:

    Declaring a correction file for a type of cassette makes it unavailable for the other cassette

    types.

    The cassette name declared in the Cassette Manager tool has to be identical to the correction file name (minus the extension *.mat), which is stored in the acquisition

    program directory, subdirectory “Config”.

    Those correction files are generated using purpose-made homogeneity standards with the help of a service tool called “BV Acquisition Tool”. (This tool is for service only and is not

    intended for a Customer use).

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    5.3 Export Log File /Audit

    A log of the instrument operation is kept and stores the following with current login &

    access level, time, and date information:

    • Acquisition start / stop (with details of the run: parameters, properties)

    • TEA Tank fill

    • Account creation / deletion / modification

    • Cassette correction creation / deletion

    • Errors

    This log can be exported by the administrator as a text file using Export Log File in the Tools menu.

    After the acquisition, properties of the run can be examined in the analysis software, and

    contain information as displayed below:

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    6 IMAGER: Maintenance

    6.1 TEA tank fill

    TEA tank fill is allowed when a set low level is reached. When the TEA level indicator goes from green to orange, a tank fill is possible.

    When the TEA level indicator is red, an acquisition cannot be started since there is not enough TEA left in the tank.

    For a TEA tank fill, you will need:

    • TEA pump TEA pump TEA pump TEA pump (When refilling for the first time, check that the input tube of the pump -going into the bottle- has the right length to go till the bottom of the bottle)

    • A new 1 liter TEA bottle A new 1 liter TEA bottle A new 1 liter TEA bottle A new 1 liter TEA bottle

    • Two 14/15 spanners (wrenches)Two 14/15 spanners (wrenches)Two 14/15 spanners (wrenches)Two 14/15 spanners (wrenches)

    • AbsorbingAbsorbingAbsorbingAbsorbing paper paper paper paper to wipe any spilled TEA

    • Safety glasses, lab coat & two pair of glovesSafety glasses, lab coat & two pair of glovesSafety glasses, lab coat & two pair of glovesSafety glasses, lab coat & two pair of gloves

    Once all these items are at hand, click on """" TEA FillTEA FillTEA FillTEA Fill " " " " in the control window. The software then depressurize the gas circuit to prepare the instrument for a TEA fill, as

    indicated in the status box :

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    Once it is over, a message box explains the following procedure :

    Procedure :Procedure :Procedure :Procedure :

    � Check the tube & pump Check the tube & pump Check the tube & pump Check the tube & pump integrityintegrityintegrityintegrity (especially underneath the fittings!) (especially underneath the fittings!) (especially underneath the fittings!) (especially underneath the fittings!) & & & & cleanlinesscleanlinesscleanlinesscleanliness.

    � Put your safety gear on.

    � Localize the filling cap of the TEA tank on the side of the β IMAGER. NOT facing NOT facing NOT facing NOT facing

    the the the the fitting, take off this cap with a 14/15 mm spanner (wrench) slowly & with a slowly & with a slowly & with a slowly & with a

    wipe underneathwipe underneathwipe underneathwipe underneath.

    � Connect the tube of the pump where the cap was.

    � Uncap the TEA bottle and push the pump body onto the bottleneck until it is tight.

    � Pump the TEA up from the bottle by squeezing the ball on top of the pump, and

    regularly check the on-screen TEA level. The display is not refreshed instantly, so

    allow for a few seconds’ delay.

    Note : After priming, lifting the bottle will result in the liquid flowing on its own into the tank. However, this proves unsafe since the bottle may be dropped & break.

    � When the bottle is empty, continue to pump to clear any TEA from the tube.

    Note : The TEA level will never reach 100 %. Do not try to fill in more than 1 liter you might damage the β IMAGER.

    � Remove the pump from the bottle (after checking there is no TEA left in the tube)

    and screw the cap back on the side of the β IMAGER. Do the same for the

    bottle’s cap.

    � You can then press the "End of Fill" button to pressurize the detector again.

    � Wash the pump under cold water, let it dry and store it until the next refill.

    Alternatively, let the pump dry in a fume cabinet.

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    Note : In case the TEA is spilled (on the floor or inside the instrument), power off the β IMAGER, wipe & wash with water or diluted acid (only the floor, not inside the machine!). See the Material Safety Data Sheet (MSDS) for TEA.

    If you do not feel comfortable with this filling procedure, do not hesitate to get in contact with our maintenance engineers to guide & assist you through a remote support link & telephone.

    Note :Note :Note :Note : Warning, Triethylamine is an organic compound of class 3, subsidiary risk of class 8, referenced n° UN or UTA: 1296. We strongly advise to read the appended MSDS for TEA. European directives for environmental rejection limits regulate Triethylamine. In normal operation, the β IMAGER is in conformity and is well below the regulations published Feb. 2d, 1998 in relation to all-type emissions.