Chapter 3Chapter 3

Biological tools Biological tools and and

InstrumentationInstrumentation

Taxonomic Taxonomic (Dichotomous) (Dichotomous)

KeyKey1a. Is the organism unicellular -go to 1a. Is the organism unicellular -go to step 2step 2

1b. Is the organism multicellular -go to 1b. Is the organism multicellular -go to step 3step 3

2a. Is the organism prokaryotic -kingdom 2a. Is the organism prokaryotic -kingdom MoneraMonera

2b. Is the organism eukaryotic -Kingdom 2b. Is the organism eukaryotic -Kingdom ProtistaProtista

Scientific MethodScientific MethodAn systematic approach to problems An systematic approach to problems

solving:solving: 1. Defining the problem1. Defining the problem

“ “ By doing Background ResearchBy doing Background Research”” 2. Formulating a hypothesis2. Formulating a hypothesis 3. Testing the hypothesis3. Testing the hypothesis 4. Recording observations4. Recording observations

5. State a conclusion5. State a conclusion

Three types of ExperimentsThree types of Experiments

1.1. ****Controlled ****Controlled Experiments**** Experiments****

2.2. Trial and Error Trial and Error

3.3. Open EndedOpen Ended

Defining the problemDefining the problem ““By doing Background By doing Background

ResearchResearch””HypothesisHypothesis – –ExperimentationExperimentation

1. 1. Control groupControl group (no change) (no change)

2. 2. Experimental groupExperimental group (make (make change)change)

control =control = variable =variable =

** State a conclusionState a conclusion

Defining the problemDefining the problem

HypothesisHypothesis – –a possible explanation for a possible explanation for events.events.

ExperimentationExperimentation (Testing the (Testing the hypothesis)hypothesis)

1. 1. Control groupControl group (no change) a source (no change) a source for for comparison comparison

2. 2. Experimental groupExperimental group (contains 2 (contains 2 variables)variables)

*Independent Variable (Manipulated) *Independent Variable (Manipulated) **Dependent Variable (responding) Dependent Variable (responding)

#’s#’sState a conclusionState a conclusion

TheoryTheory- explanation - explanation supported by experimental supported by experimental evidence.evidence.

Scientific LawScientific Law- is a - is a statement that describes statement that describes some aspect of a phenomena some aspect of a phenomena that is always true.that is always true.

MicroscopesMicroscopes1.Simple Microscopes2.Dissecting microscope3.Compound Microscope4.Phase Contrast Microscope5.Electron Microscope

• SEM• TEM

InstrumentationInstrumentationA. A. Simple microscope Simple microscope - magnifying - magnifying glassglass

1 –uses 1 lens1 –uses 1 lens B. B. Compound light microscopeCompound light microscope 1- uses two lenses1- uses two lenses a) ocular (eyepiece) 10xa) ocular (eyepiece) 10x b) objective b) objective - low power, 10x- low power, 10x - high power, 40x- high power, 40x

Compound Light Compound Light MicroscopeMicroscope

Magnifies 50x Magnifies 50x to 500x the to 500x the original sizeoriginal size

Used to look at Used to look at the whole cell the whole cell and organelles and organelles like the nucleus like the nucleus and chloroplastand chloroplast

Adjustments:Adjustments:

Coarse adjustmentCoarse adjustment – used to – used to focus under low power focus under low power

Fine adjustmentFine adjustment – used to – used to focus under high power focus under high power ONLYONLY

Used to “fine tune” the Used to “fine tune” the imageimage

Total Total MagnificationMagnification::

Magnifications: Magnifications: ocularocular = 10x = 10x

low power objective = 10xlow power objective = 10x

high power objective = 40xhigh power objective = 40x

Formula:Formula:

ocularocular x objective = total magnification x objective = total magnification

High power magnification is? High power magnification is?

1010 x 40 = 400x x 40 = 400x

Imagery:Imagery:When placing a slide under the When placing a slide under the

objective lens, the image objective lens, the image appears:appears:

-magnified-magnified-upside down-upside down-inverted (backwards).-inverted (backwards).

ExEx: e : e F F

Imagery:Imagery:(as seen under low (as seen under low

power)power)e e

F F

FF ee

Imagery:Imagery:Field of View (fov)Field of View (fov)

Scanner fov = 4000 um

F FLow fov=2000 um

What is the size of the “F”?

Imagery:Imagery:Field of View (fov)Field of View (fov)

FF 4000 um

Electron MicroscopesElectron MicroscopesAdvanced Advanced

microscope microscope magnifies 250,000x magnifies 250,000x

the original sizethe original sizeAllows for detailed Allows for detailed

observation of observation of smallsmall organellesorganelles within the cell. within the cell. Like the Like the

mitochondria and mitochondria and lysosomes.lysosomes.

Dissecting MicroscopeDissecting MicroscopeGives the viewer a Gives the viewer a

3-D image of the 3-D image of the organismorganism

Used in the Used in the dissection of small dissection of small animals and plantsanimals and plants

Low magnificationLow magnification1-10x1-10x

Other Biological ToolsOther Biological ToolsSeparation TechniquesSeparation Techniques

CentrifugeCentrifuge ChromatographyChromatography Gel electrophoresisGel electrophoresis Dialysis TubingDialysis Tubing

Dissection and micro dissection Dissection and micro dissection instrumentsinstruments

StainsStains IndicatorsIndicatorsSystem InternationalSystem International

Separation TechniquesSeparation Techniques

Centrifuge:Centrifuge: Separates of solutions based on each Separates of solutions based on each parts densityparts density

Chromatography:Chromatography: Separates Solute Separates Solute from Solvent from Solvent

Electrophoresis:Electrophoresis: separates macromolecules-separates macromolecules-either nucleic acids or proteins-on the basis of size, either nucleic acids or proteins-on the basis of size, electric charge, and other physical properties electric charge, and other physical properties

Dialysis TubingDialysis Tubing

CentrifugeCentrifuge

Separates of Separates of solutions based solutions based on each parts on each parts densitydensity

Gel ElectrophoresisGel Electrophoresis

ElectrophoresisElectrophoresis

Opposite charges at each side Opposite charges at each side causing sample to movecausing sample to moveSmaller DNA fragments move Smaller DNA fragments move faster and furtherfaster and further

IV. ElectrophoresisIV. Electrophoresis

        

       

Electrophoresis:Electrophoresis: separates nucleic acids or proteins-on the basis separates nucleic acids or proteins-on the basis

of size, electric charge, and other physical of size, electric charge, and other physical properties properties

Chromatography Chromatography Separates Solute from Solvent Separates Solute from Solvent

Distance traveled by the solute (ink)Distance traveled by the solute (ink)

Distance traveled by the solvent (liquid)Distance traveled by the solvent (liquid)Rf=Rf=

Thin-Layer Chromatography Thin-Layer Chromatography (TLC)(TLC)

ChromatographyChromatography

Retention Factor RfRetention Factor Rf

Dissection InstrumentsDissection InstrumentsTools used to perform Tools used to perform

dissection of Tissues/ dissection of Tissues/ organisms organisms

Dissecting scope Dissecting scope sometimes usedsometimes used

Micro dissection Micro dissection InstrumentsInstruments

Tools used to perform Tools used to perform dissection of individual dissection of individual cells or the removal of cell cells or the removal of cell organelles.organelles.

Most commonly used with Most commonly used with the Electron microscope the Electron microscope (greatest magnification).(greatest magnification).

Staining Staining TechniquesTechniquesThe use of solutions to make it more The use of solutions to make it more

possible for an observer to see possible for an observer to see structures of a cell easier under a structures of a cell easier under a microscopemicroscope

1. 1. IodineIodine- used primarily to stain - used primarily to stain nucleus of animal cell.nucleus of animal cell.

2. 2. Methylene blueMethylene blue- used primarily to - used primarily to view chloroplasts and the cell wall of a view chloroplasts and the cell wall of a plant cellplant cell..

Staining Staining TechniquesTechniques

IndicatorsIndicatorsDesigned to detect specific materialsDesigned to detect specific materials

INDICATORSINDICATORS DETECTS DETECTS Iodine/ Lugols solution Iodine/ Lugols solution starchstarchBenedicts solution Benedicts solution GlucoseGlucoseBiuret solutionBiuret solution proteins proteins Bromomethyl blue Bromomethyl blue carbon dioxidecarbon dioxide

and Lime waterand Lime waterLitmus paperLitmus paper acids or bases acids or bases

SI: System International, SI: System International, The Metric SystemThe Metric System

The most common system of The most common system of measurement used by scientists is measurement used by scientists is the metric system. The metric the metric system. The metric system is based on the unit 10. system is based on the unit 10. (Based on multiples of ten) The (Based on multiples of ten) The metric system has many basic units metric system has many basic units for its’ measurements. (Length = is a for its’ measurements. (Length = is a meter, etc.) meter, etc.)

SI: System International, SI: System International, The Metric SystemThe Metric System

Think of the basic units as Suffixes. Think of the basic units as Suffixes.

*Meter, Gram. Second, Liter, Degree *Meter, Gram. Second, Liter, Degree Celsius,Celsius,

Think of the abbreviations as prefixes:Think of the abbreviations as prefixes:Kilo =1000Kilo =1000Centi = 100Centi = 100thth

Milli = 1000thMilli = 1000th

ALLLL DONE!ALLLL DONE!