Abstract
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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- Ab:
-
Antibody
- ATM:
-
Ataxia Telangiectasia mutated protein kinase
- BrdU:
-
Bromodeoxyuridine
- CCD:
-
Charge-coupling device
- CGH:
-
Comparative genomic hybridization
- DF:
-
Differential fluorescence
- FCM:
-
Flow cytometry
- FIA:
-
Fluorescence image analysis
- FISH:
-
Fluorescence in situ hybridization
- FITC:
-
Fluorescein isothiocyanate
- FLICA:
-
Fluorochrome-labeled inhibitors of caspases
- FLM:
-
Fraction of labeled mitoses
- FNA:
-
Fine needle aspirate
- GFP:
-
Green Fluorescent Protein
- LSC:
-
Laser scanning cytometer
- mAb:
-
Monoclonal antibody
- MP:
-
Maximal pixel
- NF-κB:
-
Nuclear factor kappa B
- PCNA:
-
Proliferating Cell Nuclear Antigen
- PCR:
-
Polymerase chain reaction
- PI:
-
Propidium iodide
- RT-PCR:
-
Reverse transcription-polymerase chain reaction
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Supported by NCI Grant CA 28704 and by Robert A. Welke Foundation for Cancer Research.
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Pozarowski, P., Holden, E., Darzynkiewicz, Z. (2012). Laser Scanning Cytometry: Principles and Applications—An Update. In: Taatjes, D., Roth, J. (eds) Cell Imaging Techniques. Methods in Molecular Biology, vol 931. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-056-4_11
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DOI: https://doi.org/10.1007/978-1-62703-056-4_11
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