Document Detail


A standardized protocol for flow cytometric analysis of cells isolated from cerebrospinal fluid.
MedLine Citation:
PMID:  7510788     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Flow cytometry (FC) is an useful tool for the analysis of subpopulations in complex cell suspensions. When applying this method to the cerebrospinal fluid (CSF), some characteristic properties of this cell type must be taken into consideration: there are only few cells which decay rapidly in their native medium and during centrifugation. One aim of the immunostaining procedure preceding flow cytometric analysis must be to minimize cell loss in order to get an undistorted picture of 'true' CSF cell populations. Consequently, morphological flow cytometric plots of high resolution are an indispensable precondition for reliable determination of subpopulations defined by monoclonal antibody (Mab) binding. We describe a standardized protocol for the flow cytometric examination of CSF cells which minimizes undesired cell loss. By the use of a 'quality control' the extent of cell loss could be monitored. Examples of morphological flow cytometric plots are given. The subsequent determination of Mab binding subpopulations is critical when fluorescence intensities of antigen positive and negative cells are non-disjunct. A statistical test was developed for these cases often seen when cell surface determinants are expressed at low levels only.
Authors:
R Dux; A Kindler-Röhrborn; M Annas; P Faustmann; K Lennartz; C W Zimmermann
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of the neurological sciences     Volume:  121     ISSN:  0022-510X     ISO Abbreviation:  J. Neurol. Sci.     Publication Date:  1994 Jan 
Date Detail:
Created Date:  1994-04-15     Completed Date:  1994-04-15     Revised Date:  2004-11-17    
Medline Journal Info:
Nlm Unique ID:  0375403     Medline TA:  J Neurol Sci     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  74-8     Citation Subset:  IM    
Affiliation:
Department of Neurology, University Essen, Germany.
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MeSH Terms
Descriptor/Qualifier:
Antibodies, Monoclonal
Antigens, CD / analysis
Centrifugation
Cerebrospinal Fluid / cytology*
Female
Flow Cytometry / methods*
Fluorescein-5-isothiocyanate
Humans
Immunologic Techniques
Leukocyte Count
Lymphocytes / immunology
Male
Quality Control
Staining and Labeling
Time Factors
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Antigens, CD; 3326-32-7/Fluorescein-5-isothiocyanate

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