| Analysis of apoptosis by laser scanning cytometry. | |
| | |
MedLine Citation:
|
PMID: 10082299 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
Flow cytometry techniques that are widely used in studies of cell death, and particularly in the identification of apoptotic cells, generally rely on the measurement of a single characteristic biochemical or molecular attribute. These methods fail to recognize cell death lacking that attribute, as in some examples of atypical apoptosis. Since apoptosis was originally defined by morphologic criteria, we suggest that for any new cell system the cytometry-defined apoptosis be confirmed by morphologic examination. This quality assurance measure is now provided by laser scanning cytometry (LSC). LSC measurements of cell fluorescence are precise and highly sensitive, comparable to flow cytometry (FCM), and can be carried out on cells on slides, permitting cell by cell correlation of fluorescence cytometry with visual microscopic morphology. In this report we describe adaptations of various flow cytometry techniques for detection of apoptosis by laser scanning cytometry. We also describe features unique to LSC that are useful in recognizing apoptosis. Hyperchromicity of DNA, reflecting chromatin condensation, is evidenced by high maximal pixel values for fluorescence of the DNA-bound fluorochrome. Mitochondrial probes that have been adapted to LSC to measure the drop in mitochondrial transmembrane potential that occurs early in apoptosis include rhodamine 123, 3,3'-dihexiloxadicarbocyanine [DiOC6(3)], and the aggregate dye 5,5',6,6'tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide (JC-1). The changes in plasma membrane phospholipids and transport function, also early in apoptosis, are probed by a combination of the fluoresceinated annexin V and DNA fluorochromes such as propidium or 7-aminoactinomycin D. We also review methods of detection of apoptosis based on analysis of DNA fragmentation and their application to clinical oncology. Visual examination of the presumed apoptotic cells detected by cytometry makes it possible to discriminate those that are genuine from monocytes/macrophages that have ingested nuclear fragments via apoptotic bodies. Applications of flow cytometry and laser scanning cytometry in analysis of cell death are discussed and their respective advantages and disadvantages compared. |
| | |
Authors:
|
E Bedner; X Li; W Gorczyca; M R Melamed; Z Darzynkiewicz |
Related Documents
:
|
10967409 - Dose-dependent apoptosis induced by low concentrations of methylmercury in murine splen... 17240509 - Emodin induces apoptosis through caspase 3-dependent pathway in hk-2 cells. 7553879 - A triplet of nuclease proteins (np42-50) is activated in human jurkat cells undergoing ... 12738989 - Cell to cell contact required for bystander effect of the tnf-related apoptosis-inducin... 19755849 - Ghrh antagonist causes dna damage leading to p21 mediated cell cycle arrest and apoptos... 20428789 - Effects of desipramine on the cell cycle and apoptosis in ca3/7 mouse skin squamous car... 8392979 - Protein phosphatases limit tumor motility. 14973159 - Identification of multivesicular bodies as prevacuolar compartments in nicotiana tabacu... 21775119 - The effects of transformation and znt-1 silencing on zinc homeostasis in cultured cells. |
Publication Detail:
|
Type: Comparative Study; Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.; Review |
Journal Detail:
|
Title: Cytometry Volume: 35 ISSN: 0196-4763 ISO Abbreviation: Cytometry Publication Date: 1999 Mar |
Date Detail:
|
Created Date: 1999-05-25 Completed Date: 1999-05-25 Revised Date: 2013-01-16 |
Medline Journal Info:
|
Nlm Unique ID: 8102328 Medline TA: Cytometry Country: UNITED STATES |
Other Details:
|
Languages: eng Pagination: 181-95 Citation Subset: IM |
Affiliation:
|
Brander Cancer Research Institute, New York Medical College, Valhalla, USA. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Annexin A5
/
metabolism Apoptosis* Cell Membrane / metabolism Cells, Cultured DNA / analysis DNA Fragmentation Flow Cytometry Humans Image Cytometry / methods* Membrane Potentials Mitochondria / metabolism |
| Grant Support | |
ID/Acronym/Agency:
|
CA 28704/CA/NCI NIH HHS; R01 CA028704/CA/NCI NIH HHS; R01 CA028704-20/CA/NCI NIH HHS |
| Chemical | |
Reg. No./Substance:
|
0/Annexin A5; 9007-49-2/DNA |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: P2X receptor immunoreactivity in the rat cochlea, vestibular ganglion and cochlear nucleus.
Next Document: Development and validation of a computerized cytomorphometric method to assess the maturation of vag...