Laser scanning cytometry is somewhat of a hybrid technology between flow and image cytometry. The instrument is basically a fluorescent microscrope with lasers -- like an image cytometer -- but includes a motorized stage and galvonometer scanning mirror under computer control that produce flow cytometry-like signals while scanning a slide. Data is displayed in histograms and dot plots like flow cytometry. Regions can be used on the plots to select a cell population which can then be "relocated"; that is, the slide is returned to the location at which the measurement was made. In this fashion, selected cells can be viewed under bright field or epifluorescence microscopy.
From Dr. Richard Everson, annexin V/PI staining and the corresponding morphology and comets of individual cells undergoing apoptosis. First, "healthy" cells are identified as annexin V-/PI- and imaged under bright field (1) and epifluorescence (2). Necrotic and late apoptotic cells (annexin V+/PI+) are selected and similarly imaged (3, 4), as are apoptotic cells (annexin V+/PI-) in panels 6, 7, 9, and 10. After scanning, the slide was removed and subjected to the comet assay, a single cell gel electrophoresis assay to detect DNA damage. The slide was replaced on the LSC and the exact same cells were relocated for epifluorescence imaging (5, 8, and 11).
Megyeri A, Bacso Z, Shields A, Eliason JF. Development of a stereological method to measure levels of fluoropyrimidine metabolizing enzymes in tumor sections using laser scanning cytometry. Cytometry A. 64(2):62-71, 2005.
Megyari A, Kaplan J, Eliason JF. Laser Scanning Cytometry for selection of green fluorescent protein transgenic mice using small number of blood cells. J. Biochem. Biophys. Methods. 61(1-2):183-7, 2004.
Bacso Z, Eliason JF. Measurement of DNA damage associated with apoptosis by laser scanning cytometry. Cytometry. 45(3):180-6, 2001.
Bacso Z, Everson RB, Eliason JF. The DNA of annexin V-binding apoptotic cells is highly fragmented. Cancer Res. 60(16):4623-8, 2000.