Laser Microdissection and Pressure Catapulting (LMPC) Technique to Study Gene Expression in Ischemia-Reperfused Myocardium
Date & Location:
December 11, 9:15 - 10:00 a.m. in Room 16A
Presenters:
Sashwati Roy, Donald Kuhn, Chandan K. Sen
Ohio State University Medical School, Columbus, Ohio
For focal events like myocardial infarction, it is important to dissect infarction-induced biological responses as a function of space with respect to the infarct core. LMPC represents a recent variant of laser capture microdissection (LCM) that enables robot-assisted capture of catapulted tissue without direct user contact. This work represents the maiden effort to apply LCM to study spatially- resolved biological responses in myocardial infarction.
A Carl Zeiss PALM MicroBeam system with motorized precision stage and a robotic sample capturing device (RoboMover) for high throughput sample collection was employed to collect infarcted areas of the surviving ischemia-reperfused murine heart. Tissue elements captured from the infarct and non-infarct sites were collected in RNA lysis buffer followed by RNA isolation, reverse transcription and quantitative PCR. More recently, we have developed the capability to measure gene expression from myocytes and fibroblasts captured from the heart with micron-level precision.
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