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Cell Imaging Techniques pp 187–212Cite as

Laser Scanning Cytometry: Principles and Applications—An Update

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 931))

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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Abbreviations

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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Acknowledgement

Supported by NCI Grant CA 28704 and by Robert A. Welke Foundation for Cancer Research.

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Authors and Affiliations

  1. The Brander Cancer Research Institute, New York Medical College, Valhalla, NY, USA

    Piotr Pozarowski & Zbigniew Darzynkiewicz

  2. Department of Clinical Immunology, School of Medicine, Lublin, Poland

    Piotr Pozarowski

  3. CompuCyte Corporation, Westwood, MA, USA

    Elena Holden

Authors
  1. Piotr Pozarowski
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  2. Elena Holden
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  3. Zbigniew Darzynkiewicz
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Corresponding author

Correspondence to Zbigniew Darzynkiewicz .

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Editors and Affiliations

  1. College of Medicine, Professor, Department of Pathology, University of Vermont, Beaumont Ave 89, Burlington, 05405-1742, Vermont, USA

    Douglas J. Taatjes

  2. , Abt. Zell- und Molekularpathologie, Universität Zürich, Schmelzbergstr. 12, Zürich, 8091, Switzerland

    Jürgen Roth

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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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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-055-7

  • Online ISBN: 978-1-62703-056-4

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