CONTACT technology of Laser Capture Microdissection (PixCell II, Arcturus)
Principle and main steps of LCM (Background)
LCM uses a heat-generating infrared laser to fuse a temperature-sensitive transfer membrane with the underlying histological tissue. The membrane is mounted onto a special microfuge cap and detachment of the membrane results in only selected specimen adhering to the transfer membrane (positive cell sorting).
- CHOOSE SAMPLE
Paraffin embedded tissue for Þ DNA-extraction Frozen (fresh) tissue or smear Þ for RNA, DNA or protein extraction
- PREPARE
Follow routine protocols with minor changes for preparing a tissue or smear on a standard slide (all protocols available upon request). Apply a Prep Strip to flatten tissue and remove loose debris prior to LCM
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- LOCATE
Visualize the sample trough the video monitor or the microscope. Locate the cell(s) of interest and position the CapSure with the transfer film (plastic cap, d=0.6 cm) over the cells.
- CAPTURE
Pulse the low power infrared laser to activate the transfer film. The cell(s) within the laser beam adhere to the transfer film.
- MICRODISSECT
Lift the cap with the desired cell(s) attached to the film surface. The surrounding tissue remains intact.
- INCUBATE
Pipette digestion buffer directly into the microcentrifuge tube. Place the cap directly onto a standard microcentrifuge tube containing the extraction buffer using assembling device. Invert the microcentrifuge tube and incubate according to extraction procedures.
- ANALYZE
Centrifuge to collect cell extract at the bottom of the tube. The cell contents (DNA, RNA or protein) are ready for molecular analysis.
