Evaluation of scanning cytometer fluorometry performance

Susanne Heynen; Jeffrey H. Price M.D.

doi:10.1117/12.349205

1 June 1999 Evaluation of scanning cytometer fluorometry performance

Susanne Heynen, Jeffrey H. Price M.D.

Author Affiliations +

Proceedings Volume 3604, Optical Diagnostics of Living Cells II; (1999) https://doi.org/10.1117/12.349205
Event: BiOS '99 International Biomedical Optics Symposium, 1999, San Jose, CA, United States

Abstract

Recent advances in autofocus, image segmentation, and fluorescence lamp stabilization have resulted in a fully automated high-speed image cytometer. This system performs computer-controlled autofocus, incremental stage scanning, image processing and data storage at a combined 1 Hz field rate. DAPI stained 3T3 cells that were cultured on microscope cover slips were scanned using the scanning cytometer. The integrated intensity for each detected cell object was computed and stored as well as its area and information about its location. Performance of the system was evaluated by repeatedly scanning cellular objects and comparing the results of each object over several hundred scans. This method of performance evaluation has the advantage of being independent of variations in stain homogeneity of the slide being scanned. For several hundred repeat scans, coefficients of variation (CVs) of significantly less than 1% were consistently obtained for the integrated intensity of each cellular object in the experiment.

Citation Download Citation

Susanne Heynen and Jeffrey H. Price M.D. "Evaluation of scanning cytometer fluorometry performance", Proc. SPIE 3604, Optical Diagnostics of Living Cells II, (1 June 1999); https://doi.org/10.1117/12.349205

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