摘要:

After reviewing basic technical considerations, we discuss some applications of flow cytometry in French laboratories. This methodology is used in several areas: oncology, cellular pharmacotoxicology, molecular biology and genetics, immunology, as well as cellular biology and physiology. We also examine the evolution of this technique in two directions: on the one hand, the appearance of increasingly sophisticated instruments; on the other, the development of less expensive and less complicated apparatuses principally directed at clinical applications.

ISSN:0248-4900    

DOI:10.1111/j.1768-322X.1986.tb00492.x

出版商:Blackwell Publishing Ltd    

年代:1986

数据来源: WILEY

摘要:

Flow cytometry has been extensively used to provide accurate estimates of the relative amounts of various cellular constituents (DNA, RNA, proteins) for cell kinetic studies. Multiparametric analysis also supports the recent concept that cell growth and the DNA division cycle may be under distinct regulatory mechanisms. Moreover, metabolic subcompartments of the cell cycle, distinguished by flow cytometry, have offered a highly sensitive cell classification in comparison with the conventional distinction of the four main phases of the cell cycle. Finally, a new sensitive and powerful technology, BrdU/DNA analysis, represents a remarkable maturing of a very useful alternative for the study of DNA synthesis and cell cycle traverse.

ISSN:0248-4900    

DOI:10.1111/j.1768-322X.1986.tb00493.x

出版商:Blackwell Publishing Ltd    

年代:1986

数据来源: WILEY

摘要:

Flow DNA analysis was performed on samples from 71 surgically removed lung cancers and from 145 patients undergoing bronchoscopy. Abnormal DNA stem lines, characterized by their DNA index (DI), were frequently observed in operated lung carcinomas (87%). Two or three abnormal DNA stem lines were discovered simultaneously in 10% of the samples. The mean DI of all abnormal tumor stem lines was lowest for rare tumor cell types and highest for adenocarcinomas. Intermediate mean DI values were found for epidermoid and small cell carcinomas, which were among the most proliferative tumors. The high rate of false negative results suggests poor diagnostic reliability of flow DNA analysis on bronchoscopic samples. However, the method appears to provide a promising objective tool capable of evaluating tumor behavior and prognosis.

ISSN:0248-4900    

DOI:10.1111/j.1768-322X.1986.tb00494.x

出版商:Blackwell Publishing Ltd    

年代:1986

数据来源: WILEY

摘要:

Microspheres of latex ranging from 0.1 to 10 microns in diameter are nowadays commercially available. Labelled with different fluorochromes, they are good standards to check the optical path and to calibrate flow cytometers. The kits available contain microspheres highly selected with a precise size and a narrow distribution, so as to allow an easy alignment of instruments. Mixed with the cell preparations, microspheres may be used as an internal standard, in order to compare samples from day to day. Phagocytic activity has been investigated after incubation of cells with fluorescent microspheres. Covalently bound to ligands such as antibodies, microspheres become a powerful and specific reagent to label cell surface antigens. The positive signal is enhanced, allowing the detection of poorly represented epitopes. This paper reviews some examples of these applications published in the flow cytometry field.

ISSN:0248-4900    

DOI:10.1111/j.1768-322X.1986.tb00495.x

出版商:Blackwell Publishing Ltd    

年代:1986

数据来源: WILEY

摘要:

Cellular pharmacology is defined as the study of drug effects on various cell functions. Flow cytometry enriches cellular pharmacology by the following possibilities for efficient analysis. Firstly, the determination of toxic concentrations can be approached by the assessment of cell viability. However, due to the existence of many fluorescent DNA probes, most studies are devoted to the investigation of products acting on cell division, particularly in the area of antineoplastic drugs. The effects of drugs on respiration can be approached by analysis of mitochondrial activities. On the other hand, the studies of drug actions on cell differentiation functions have been started using antisera or monoclonal antibodies to cell‐specific proteins such as collagen and keratin. Flow cytometry appears to be more and more important in the progress of cellular toxicology and pharmacolog

ISSN:0248-4900    

DOI:10.1111/j.1768-322X.1986.tb00496.x

出版商:Blackwell Publishing Ltd    

年代:1986

数据来源: WILEY

摘要:

Almost all antineoplastic drugs are able to delay—or block—cells in a particular phase of the cell cycle. Few clinically active drugs seem to interact with the G1‐states where cell growth can be arrested, although new compounds could be of interest with this respect. In contrast, most antineoplastic agents interact with DNA and/or DNA metabolism and have been shown to provoke a delay in G2. This could be the consequence of the DNA damage or of interference with controls which take place within the G2

ISSN:0248-4900    

DOI:10.1111/j.1768-322X.1986.tb00497.x

出版商:Blackwell Publishing Ltd    

年代:1986

数据来源: WILEY

摘要:

The effects of individual drugs on cell cycle progression can often be determined by analyzing the DNA distribution of cultured cells at appropriate times after drug administration. In addition, to cell counts, RNA and/or protein content, the alterations in cell cycle distribution of drug‐treated cells can yield information on the potential cell cycle phase specificity of the drug. However single parameter DNA analysis, as currently used, can give inaccurate information when drug effects are associated with cytokinesis perturbations, in spite of various statistical and mathematical analyses. Scanning flow cytometry could be an interesting alternative for the detection of binucleate cell

ISSN:0248-4900    

DOI:10.1111/j.1768-322X.1986.tb00498.x

出版商:Blackwell Publishing Ltd    

年代:1986

数据来源: WILEY

摘要:

Improvement in our knowledge in cellular biology is largely related to the use of new tools in quantitative cytology. Among them, flow cytometry was developed with numerous applications in the field of immunology including fundamental and applied research. Since its early beginning it has been associated with monoclonal antibodies to identify immuno‐competent cells, to quantify changes in expression of surface determinants, to separate cells subsets prior to the test of their functional properties. Major advances gained using either single or dual‐laser systems, multicolour fluorescence and computer facilities for multi‐parametric analysis. Using this methodology it was possible to correlate analysis of cell cycle phases and membrane antigens expression. Applications have been developed for the analysis of new drugs in vitro, the evaluation of immunomodulating treatment and for clinical investiga

ISSN:0248-4900    

DOI:10.1111/j.1768-322X.1986.tb00499.x

出版商:Blackwell Publishing Ltd    

年代:1986

数据来源: WILEY

摘要:

Although flow cytometry is not yet widely used for diagnostic purposes, it provides a powerful tool for investigating structural and functional properties of immune‐responsive cells. By far, the largest application of immunofluorescence is in identifying specific cell surface features thereby allowing the discrimination of lymphocyte subpopulations (phenotyping normal and malignant cells), for analysing complex receptors on neutrophils and monocytes, for detecting the appearance of new surface antigens induced by cell differentiation or activation. Besides this, the technique sets objective standards for appreciating cell activation (changes in RNA content, increased phagocytosis, DNA synthesis, changes in membrane fluidity, pH gradient changes). Computer processing of measurements allows rapid accurate studies of several properties simultaneousl

ISSN:0248-4900    

DOI:10.1111/j.1768-322X.1986.tb00500.x

出版商:Blackwell Publishing Ltd    

年代:1986

数据来源: WILEY

摘要:

The plasma membrane of enterocytes comprises two structurally and functionally distinct domains. These are the apical brush border, containing digestive hydrolases and glycocalyx, and the basolateral domain, characterized by other specific markers. Using a fast and easy subcellular fractionation, we purified four membrane vesicle fractions from rabbit small intestinal mucosa: brush border, basolateral, rough endoplasmic reticulum and Golgi + smooth endoplasmic reticulum. Using flow cytometry, the fluorescence polarization of diphenylhexatriene was determined in brush border and in basolateral + Golgi + smooth endoplasmic reticulum membrane fractions in order to investigate changes in the membrane fluidity of both fractions and to compare the results obtained with those of spectroscopic techniques. Moreover, it was possible with flow cytometry to detect and quantify basolateral and brush border markers by using polyclonal and monoclonal antibodies. The advantages of flow cytometry in the detection of brush border membrane markers found in small amounts in the basolateral domain are discussed. Finally, flow cytometry holds great promise for the analysis and sorting of subcellular fractions.

ISSN:0248-4900    

DOI:10.1111/j.1768-322X.1986.tb00501.x

出版商:Blackwell Publishing Ltd    

年代:1986

数据来源: WILEY