Multiparameter analysis of progeny of individual cells by laser scanning cytometry

Bedner, Elżbieta; Ruan, Qiongfang; Chen, Sophie; Kamentsky, Louis A.; Darżynkiewicz, Zbigniew

doi:10.1002/1097-0320(20000801)40:4<271::aid-cyto3>3.0.co;2-c

Cited by 18 publications

(17 citation statements)

References 23 publications

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“…Thus, mechanistic studies can be carried out using LSC, for example to study the difference between the aneugen-and clastogen-induced MN, or to study whether different clastogen types preferentially induce a particular chromosome or set of chromosomes to separate from the rest of the chromatin and form a single or multiple MN per cell. One has to mention that in analogy to the present application, the FISH capabilities of LSC were extended before to measure DNA content of individual cell nuclei within cell colonies, in the clonogenicity assay (27).…”

Section: Discussionmentioning

confidence: 99%

“…In the second approach the threshold contour was set on the data from the sensor measuring FITC fluorescence and the CompuCyte software designed to measure fluorescence in situ hybridization (FISH) spots (21-23) was used. This approach, which was similar to that used by us before to measure individual cell colonies (27) allowed us to count MN and nuclei and measure intensity of their green and red fluorescence, in each individual cell. The ratio of green to red (integral) fluorescence intensities as well as the ratio of circumference to area of the measured nuclei and MN also were recorded.…”

Section: Fluorescence Measurementsmentioning

confidence: 99%

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Micronuclei assay by laser scanning cytometry

et al. 2001

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Section: Discussionmentioning

confidence: 99%

Section: Fluorescence Measurementsmentioning

confidence: 99%

Micronuclei assay by laser scanning cytometry

et al. 2001

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“…Another application of LSC utilizing the FISH approach was demonstrated in the analysis of progeny (clones) of individual cells (56). In this application, cellular protein and DNA were stained with fluorochromes of different color while the product of tumor suppressor gene p53 or estrogen receptor was detected immunocytochemically, with still another color fluorescent dye.…”

Section: Applications Of Lsc Utilizing the Software Designed For Fmentioning

confidence: 99%

Laser Scanning Cytometry: Principles and Applications—An Update

Pożarowski

Holden²,

Darżynkiewicz

2012

Methods in Molecular Biology

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Laser scanning cytometer (LSC) is the microscope-based cytofluorometer that offers a plethora of unique analytical capabilities, not provided by flow cytometry (FCM). This review describes attributes of LSC and covers its numerous applications derived from plentitude of the parameters that can be measured. Among many LSC applications the following are emphasized: (a) assessment of chromatin condensation to identify mitotic, apoptotic cells, or senescent cells; (b) detection of nuclear or mitochondrial translocation of critical factors such as NF-κB, p53, or Bax; (c) semi-automatic scoring of micronuclei in mutagenicity assays; (d) analysis of fluorescence in situ hybridization (FISH) and use of the FISH analysis attribute to measure other punctuate fluorescence patterns such as γH2AX foci or receptor clustering; (e) enumeration and morphometry of nucleoli and other cell organelles; (f) analysis of progeny of individual cells in clonogenicity assay; (g) cell immunophenotyping; (h) imaging, visual examination, or sequential analysis using different probes of the same cells upon their relocation; (i) in situ enzyme kinetics, drug uptake, and other time-resolved processes; (j) analysis of tissue section architecture using fluorescent and chromogenic probes; (k) application for hypocellular samples (needle aspirate, spinal fluid, etc.); and (l) other clinical applications. Advantages and limitations of LSC are discussed and compared with FCM.

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“…Possible clinical applications of this approach could combine live cell function assays in vitro such as oxidative burst or phagocytosis with immunophenotyping after fixation. At the other end of the spectrum are cell clonogenicity (85) and comet assays (20,21,86) performed by LSC. In cardiology, the analysis of platelets and platelet-endothelial interactions are of particular importance for the prediction of cardiac events; methods have been adapted for analysis by LSC (87,88).…”

Section: Other Applicationsmentioning

confidence: 99%