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Laser Microdissection Analysis Platform

Laser Microdissection Analysis Platform. Administrative Coordination : M. Nakache (CR1) Scientific Coordination: T. Jay (DR2 ) & L. Villanueva (DR2) Experimental Coordination: E. Davenas (IR1). Laser Microdissection : a tool within a process. (2). Genome Transcriptome

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Laser Microdissection Analysis Platform

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  1. Laser Microdissection Analysis Platform Administrative Coordination: M. Nakache (CR1) Scientific Coordination: T. Jay (DR2) & L. Villanueva (DR2) Experimental Coordination: E. Davenas (IR1)

  2. Laser Microdissection : a toolwithin a process (2) Genome Transcriptome PCR RT-PCR Proteome Western blot 2D-gel, 2D-DIGE Mass Spectrometry Extraction (1) Tissue Prelevement (4) localisation Histologic Observations (3) Histochemical-immunity In Situ Hybridation TUNEL Laser Microdissection

  3. Goals Cell or tissu isolation under morphologic control in order to perform molecular biology studiesfrom either frozen or paraffin-embedded tissue slices or cell cultures

  4. Fromtissue to molecule (1) 1.Sample preparation and cutting - fixation - paraffin embedding - freezing 2.Visualisation of areas of interest - staining - fluorescence 3.Selective Microdissection - no contact, no contamination - anysize (µm2 -> mm2), shape, thickness(< 150 µm)

  5. LMPC PALM® SystemLaser Microdissection & Pressure Catapulting The laser beamcutsthe area of interest then, by adjustingitsenergy and itsfocusing, itcatapultsthe selectedregionintothe cap of a tube placedjustover. This cap isfilledwitha lysis buffer adequate for subsequentanalysis: - DNA - RNA - Protein

  6. Fromtissue to molecule (2) 4.Extraction and preparation of specific molecules: DNA, RNA, proteins 5. Analysis of pertinent, reproducibleand specificresults

  7. Molecular analysis: DNA, RNA, Proteins DNA • DNA Extraction • Tumor Analysis • DNA Fingerprinting • RNA Extraction • Quantitative RT-PCR • Microarray analysis RNA • HPLC • SDS-PAGE • MassSpectroscopyanalysis (MALDI, SELDI) Protein

  8. Materialsavailable PALM®Microbeam Laser Microdissector(Zeiss) Cryostat HM550(MM France) Microtome HM450(MM France) UV CrosslinkerBLX-254 (BIO-LINK) Hood Chemfree90S(Oxygen)

  9. Laser Microdissection GlobusPallidus Subthalamic Nucleus Black Matter 2,5X 2,5X 2,5X 10X 10X 10X

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