The Synergy of Selective Laser Microdissectionand Sensitive AnalysisLeica Microsystems – QIAGEN Application Support

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Sectioning Staining Microdissection Isolation Analysis

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Sectioning Staining

Reliable instruments for infiltration,embedding and sectioningPrepare tissue sections by using a variety of sec-tioning techniques. Choose the appropriate Leicainstrument for your application: cut frozen tissuewith a cryostat, fixed and paraffin-embedded tis-sues using the Leica RM22 family of rotary microto-mes, or native, unfrozen tissue under physiologicalconditions with a Leica VT1200/1200 S vibratingmicrotome.

Efficient staining of tissue sectionsStain your fixed sections and visualize cell structures athigh resolutions. Staining workstations from Leica allowseveral staining protocols to run simultaneously, and canbe programmed to satisfy customized requirements. Built-in filters reduce exposure to hazardous fumes, and thestaining process can be monitored through a transparentlid.

Microdissection

Fast, selective and easy to use!Collect selective dissected single cell(s) and tissueregions of any shape or size. The Leica LMD6500/7000are fully automated laser microdissection systems enab-ling the user to collect easily ultra-pure samples with aprecise, contamination-free method-simply by gravity.The unique systems allow you to harvest directly one ormore cell types from a slide or a petri dish. Excise selec-tive cells or metaphase chromosomes from a wide rangeof samples, including paraffin-embedded sections, fro-zen sections, blood smears, and stained or immuno-labe-led specimens.

Leica RM2265 –Rotary Microtome, highperformance motorizedand manual sectioning.

Leica CM1950 – Versatileresearch cryostat providesexcellent frozen sectioningquality and reproducibili-ty, especially for largespecimens such as braintissue.

Leica LMD7000 – Experience a new levelof speed.

Leica ST5020 –Automated stainingfor highly efficientworkflow

Frame foil slides andglass foil slides

Leica VT1200/1200 S –Vibrating blade microtome toprepare high-quality fixed andunfixed sections, ideal for neu-roscience research.

Paraffin section unstained (intestines) HE stained tissue section (intestines) After microdissection/dissectate in PCR tube

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Leica ST4020 –Quick and easyautomated stainingof frozen sections

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Isolation of Analytes

Efficient purification of DNA, RNA, miRNA,and proteinsPurify analytes from laser-microdissected tissue sectionsusing QIAGEN® Sample Technologies that are special-ly designed to process very small amounts of startingmaterial. Efficient purification of DNA and RNA fromlaser-microdissected fresh-frozen tissues is achievedusing the QIAamp® DNA Micro Kit (cat. no. 56304)and the RNeasy® Plus Micro Kit (cat. no. 74034), respec-tively. For laser-microdissected formalin-fixed, paraffin-embedded (FFPE) tissues, the QIAamp DNA FFPE TissueKit (cat. no. 56404), RNeasy FFPE Kit (cat. no. 73504),and miRNeasy FFPE Kit (cat. no. 217504) maximizerecovery of usable DNA, RNA, and miRNA, respective-ly. Simultaneous purification of genomic DNA and totalRNA (including small RNA) from FFPE tissue sections orlaser-microdissected cryosections is made possible withthe AllPrep® DNA/RNA FFPE Kit (cat. no. 80234) or theAllPrep DNA/RNA Micro Kit (cat. no. 80284), respecti-vely. Full-length proteins can also be extracted fromwhole FFPE tissue sections using the Qproteome® FFPETissue Kit (cat. no. 37623).

Molecular Analysis

Highly specific and sensitive amplification fordetailed characterization of nucleic acidsAmplify nucleic acids using QIAGEN Assay Technologies,which are ideally suited for very small amounts of startingmaterial. Highly specific hot-start PCR solutions such asHotStarTaq® Plus DNA Polymerase (cat. no. 203601) andthe QIAGEN Multiplex PCR Plus Kit (cat. no. 206152) faci-litate fast and successful molecular analysis of DNA. TheREPLI-g® series enables whole genome amplification(www.qiagen.com/PG/REPLI-g), and the QIAGENOneStep RT-PCR Kit (cat. no. 210210) allows one-step RT-PCR of any RNA template without optimization, and withmaximum sensitivity, making them highly suited for appli-cations including viral load determination. For gene expres-sion analysis, the QuantiFast® Probe RT-PCR Plus Kit (cat.no. 204482) is highly suited for sensitive one-step quantifi-cation of RNA targets, including from FFPE samples con-taining highly degraded RNA. Combined with genomewi-de, predesigned QuantiFast Probe Assays, they deliverguaranteed results in probe-based real-time PCR detection.For two-step qRT-PCR, the QuantiTect® ReverseTranscription Kit (cat. no. 205311) is optimized for cDNAsynthesis with integrated genomic DNA removal from verysmall amounts of RNA (10 pg to 1 μg). Quantitative PCRand RT-PCR using QuantiFast Multiplex PCR and RT-PCRKits (cat. nos. 204543 and 204643) allow analysis of mul-tiple targets (2–4) in 1 reaction vessel. For microRNA rese-arch, the miScript PCR Starter Kit (cat. no. 218193) enablesquantification of mature and precursor miRNA. QIAGEN´sPCR kits, together with automated platforms for PCR setupand DNA fragment analysis, ensure reproducibility and con-venience, along with fast and consistent delivery of success-ful results.

Achieve sensitive and specific resultsin conventional and real-time amplifi-cation.

Pure nucleic acid

Sensitive analysisFrozen Formalin

M tissue fixed tissue M

500 bp

QIAGEN kits provide high-qualityRNA and DNA from laser microdis-sected specimens

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Convenient Kits

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O n e S o l u t i o n – Tw o I n n o v a t i v e P a r t n e r sThe Leica–QIAGEN AgreementLeica is a leading supplier of high precision optical microscopes and related instruments, and provides efficient tools for all the steps required for laser microdissection of tissue samples.QIAGEN has a worldwide reputation for its technologies for nucleic acid purification, and provides optimized reagents and protocols for purification and analysis of nucleic acids fromdissected samples. The agreement between QIAGEN and Leica combines the expertise of two leading companies to bring you complete solutions for analyzing nucleic acids from laser-microdissected specimens.

For detailed information contact your local partner!

Leica Microsystems CMS GmbHErnst-Leitz-Straße 17 – 37D-35578 Wetzlar, GermanyTel. +49 (0) 64 41-29 0Fax +49 (0) 64 41-29 25 99www.leica-microsystems.com

QIAGEN GmbHQIAGEN Strasse 140724 Hilden,GermanyTel. +49 (0) 2103-29-12400Fax +49 (0) 2103-29-22000www.qiagen.com

Leica Microsystems – an international company with a strong network of customer servicesAustralia: North Ryde Tel. +61 2 8870 3500 Fax +61 2 9878 1055

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For up-to-date licensing information and product-specific disclaimers, see the respective QIAGEN kit handbook oruser manual. QIAGEN kit handbooks and user manuals are available at www.qiagen.com or can be requestedfrom QIAGEN Technical Services or your local distributor.

Trademarks: QIAGEN®, QIAamp®, AllPrep®, HotStarTaq®, Qproteome®, QuantiFast®, QuantiTect®, REPLI-g®,RNeasy®(QIAGEN Group).

QIAGEN — a wide range of products and services for analyte isolation and analysis

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