Laser scanning cytometry: principles and applications.
The laser scanning cytometer (LSC) is the microscope-based cytofluorometer that offers a plethora of analytical capabilities. Multilaser-excited fluorescence emitted from individual cells is measured at several wavelength ranges, rapidly (up to 5000 cells/min), with high sensitivity and accuracy. The following applications of LSC are reviewed: (1) identification of cells that differ in degree of chromatin condensation (e.g., mitotic or apoptotic cells or lymphocytes vs granulocytes vs monocytes); (2) detection of translocation between cytoplasm vs nucleus or nucleoplasm vs nucleolus of regulatory molecules such as NF-kappaB, p53, or Bax; (3) semiautomatic scoring of micronuclei in mutagenicity assays; (4) analysis of fluorescence in situ hybridization; (5) enumeration and morphometry of nucleoli; (6) analysis of phenotype of progeny of individual cells in clonogenicity assay; (7) cell immunophenotyping; (8) visual examination, imaging, or sequential analysis of the cells measured earlier upon their relocation, using different probes; (9) in situ enzyme kinetics and other time-resolved processes; (10) analysis of tissue section architecture; (11) application for hypocellular samples (needle aspirate, spinal fluid, etc.); (12) other clinical applications. Advantages and limitations of LSC are discussed and compared with flow cytometry.
Pozarowski
Piotr
P
The Brander Cancer Research Institute, New York Medical College, Valhalla, NY, USA.
Holden
Elena
E
Darzynkiewicz
Zbigniew
Z
D002467:Cell Nucleus / Q000737:chemistry / Q000378:metabolism; D002478:Cells, Cultured; D003593:Cytoplasm / Q000737:chemistry / Q000378:metabolism; D006098:Granulocytes / Q000166:cytology; D006801:Humans; D016130:Immunophenotyping; D047569:Laser Scanning Cytometry / Q000295:instrumentation / Q000379:methods; D008214:Lymphocytes / Q000166:cytology; D015162:Micronucleus Tests; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D009000:Monocytes / Q000166:cytology; D012984:Software