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Proceedings Paper

Laser ablation cell sorting in scanning cytometry
Author(s): Feimo Shen; Jeffrey H. Price
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Paper Abstract

Flow cytometry has been an important tool for automated cells sorting. However, the lack of good sensitivity prevents it from being used for rare events sorting; furthermore, fragile cells, anchorage-dependent cells, and clump forming cells cannot be sorted this way. A fully automated, high-speed scanning cytometer with autofocus and image segmentation is capable of accurately locating contaminant cells in a monolayer cell population. A laser ablation system was incorporated into the cytometer to negatively sort out the unwanted cells by applying a focused, ultra-short laser pulse (sub-micron diameter, pulse duration = 4 nsec, wavelength - 500 nm) to each targeted cell. Due to the high power density (approximately 1010 W/cm2) that was present at the focal point, disruptive mechanical forces were generated and were responsible for the kill. Fluorescently stained NIH-3T3 fibroblast cells were used as a model contaminant target ells in an unstained NIH-3T3 population to determine the identification-kill effectiveness. The contaminant cells were stained with the fluorochrome CellTracker Blue CMAC, whereas the background cells were left intact. Ablation pulses were applied in frame-by-frame increment batches to the cell culture on the microscope. The negative sorting effectiveness was analyzed by automatically re-scanning the post-ablation cell culture in phase contrast and propidium iodide stained epi fluorescent fields to verify cell death.

Paper Details

Date Published: 10 May 2001
PDF: 11 pages
Proc. SPIE 4260, Optical Diagnostics of Living Cells IV, (10 May 2001); doi: 10.1117/12.426777
Show Author Affiliations
Feimo Shen, Univ. of California/San Diego (United States)
Jeffrey H. Price, Univ. of California/San Diego (United States)


Published in SPIE Proceedings Vol. 4260:
Optical Diagnostics of Living Cells IV
Daniel L. Farkas; Robert C. Leif; Robert C. Leif, Editor(s)

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