摘要:

Summary—Because flow cytometry permits the analysis of individual whole cells, one of the key requirements in selecting a probe is its ability to target the site of interest into cells. In addition, dyes must possess ideal properties (ie extinction coefficient, Stoke's shift) rendering them appropriate for this methodology. Other characteristics, such as fluorescence quenching and energy transfer, inherent to the staining, provide numerous applications in flow cytometry. The available fluorophores used in flow cytometry are classified according to their cellular incorporation and binding. Thus, probes are presented and discussed as follows: 1) dyes of cellular components (DNA, RNA, proteins, lipids); 2) probes of membrane potential; 3) fluorophores that are sensitive to their microenvironment (pH, calcium, etc); and 4) those used for measurement of enzymatic activities into cell

ISSN:0248-4900    

DOI:10.1016/0248-4900(93)90109-R

出版商:Blackwell Publishing Ltd    

年代:1993

数据来源: WILEY

摘要:

Summary—Numerous flow cytometric analyses are based on DNA content studies. We have considered firstly monoparametric cell cycle analyses, which only take DNA content into account, but are sometimes of limited interest. Then, we have presented multiparametric analyses, which can be used to improve cycle phase identification by taking simultaneously into account DNA and other cellular components, or by considering some events occurring during cell cycle. Finally, we have discussed monoparametric and multiparametric cell cycle analysis interest in various application fields, particularly in pharmacology, toxicology, tumoral pathology and higher plant system studie

ISSN:0248-4900    

DOI:10.1016/0248-4900(93)90110-Z

出版商:Blackwell Publishing Ltd    

年代:1993

数据来源: WILEY

摘要:

Summary—RNA was less studied by flow cytometry (FCM) in comparison to other cellular components. Moreover, most of these studies were applied to the measurement of total RNA. Nevertheless, such a quantification, essentially by the use of acridine orange, but also other dyes or dye combinations, permitted the acquisition of important information about the general metabolism, and the characterization of different tumoral cell types. More recently, FCM has been used to quantify specific mRNA, in two different approaches: the first one employes the sorting followed by RNA extraction and/or detection by specific radiolabeled probes; in the second approach, the use of fluorescencein situhybridization (FISH) on cells in suspension allowed the direct quantification of specific mRNA by FC

ISSN:0248-4900    

DOI:10.1016/0248-4900(93)90111-Q

出版商:Blackwell Publishing Ltd    

年代:1993

数据来源: WILEY

摘要:

Summary—Flow cytogenetic is widely used since 1975, and essentially contributes to caryotype analysis and chromosome sorting. The principles of experimentation and its possibilities and limitations are now well known. Recently several new technologies have appeared. What attitude should the cytometrist adopt regarding PCR, microdissection of chromosomes,in situhybridization, slit‐scan flow cytometry or image analy

ISSN:0248-4900    

DOI:10.1016/0248-4900(93)90112-R

出版商:Blackwell Publishing Ltd    

年代:1993

数据来源: WILEY

摘要:

Summary—An introduction is given to the literature concerning methods and objectives for cytometric DNA analysis of plant nuclei. This area has gained relevance with applications in plant breeding and seed production industries, where laboratories unfamiliar with cytometry are adopting the method. An extensive graphical guide to interpreting DNA histograms and their problems is given. Conversely, cytometry laboratories unfamiliar with plant sciences will find herein a guide, and references, to adapt their methods to plant material. A table of 2C values reassessed by flow cytometry for 70 plant species, plus the genome composition (GC%) in many instances, is also include

ISSN:0248-4900    

DOI:10.1016/0248-4900(93)90113-S

出版商:Blackwell Publishing Ltd    

年代:1993

数据来源: WILEY

摘要:

Summary—Flow cytometric studies of spermatogenesis have been advanced by the need for: i) rapid, sensitive, objective and multiparameter measurements of reproductive effects due to environmental, occupational, and therapeutic exposure to toxicants; and ii) assessment of fertility potential of human and animal sperm. As a consequence, various flow cytometric techniques are already available to identify germ cell subpopulations undergoing both proliferative and maturative processes in normal and perturbed conditions. Significant improvements have been introduced in order to investigate the spermatogenic complex differentiation pathway and the apparent uniformity of mature sperm. Flow cytometry (FCM) has been applied to the measurement of both testis and sperm cells in a variety of species, including man. End points considered in toxicology studies are: altered testicular germ cell ratios, DNA and RNA content, increase of the coefficient of variation, induction of diploid elongated spermatids and diploid sperm, altered nuclear morphology, sperm cell viability, mitochondrial function and sperm chromatin structure. Precise DNA content measurements allow accurate analysis to determine the proportion of X‐ and Y‐chromosome bearing sperm and sorting of these subpopulations for gender preselection. FCM technology has reached a maturation level that allows its inclusion in the list of available and routine methods for reproductive studies in human and animal popula

ISSN:0248-4900    

DOI:10.1016/0248-4900(93)90114-T

出版商:Blackwell Publishing Ltd    

年代:1993

数据来源: WILEY

摘要:

Summary—Flow cytometry has recently become a very important tool in the study of the mechanisms of multidrug resistance (MDR). This technique allows rapid access to information regarding two characteristics related to the MDR phenotype: i) the level of overexpression of the P‐glycoprotein; and ii) its functional aspect in expulsion of the cytotoxic agents to which the cell is exposed. In pharmacology, flow cytometry also allows evaluation of the capacity of modulating agents to reverse this resistance, and contributes a deeper insight into the mechanism of action of new molecu

ISSN:0248-4900    

DOI:10.1016/0248-4900(93)90115-U

出版商:Blackwell Publishing Ltd    

年代:1993

数据来源: WILEY

摘要:

Summary—Flow cytometry (FCM) analysis of cellular DNA content has become an increasingly important clinical research tool for the measurement and identification of abnormal cell populations. Examination of cellular DNA content can provide information for use in cell cycle analysis in cancer studies. Most researchers would agree that tumor cell proliferation indices should be leading candidates in the search for prognostic factors in breast cancer. FCM is certainly a powerful technique but the assignment of DNA index and synthetic phase of cycle are subject to a number of technical pitfalls which limit the accuracy and reproducibility of this information. This paper will focus on the difficulties encountered in FCM‐DNA analyses performed in a clinical cont

ISSN:0248-4900    

DOI:10.1016/0248-4900(93)90116-V

出版商:Blackwell Publishing Ltd    

年代:1993

数据来源: WILEY

摘要:

Summary—Rapid developments in computer technology combined with new fluorescent dyes and monoclonal antibody production have led to the development of a powerful tool: flow cytometry. The techniques of flow cytometry can be applied in a wide clinical field, from routine tasks to research in immunology and hematology. The availability of automated instruments and standardized sample preparation methods have led to its daily clinical use, helping diagnosis, prognosis or monitoring therapy. Further advances will be made with the introduction of multi‐parametric analysisiequantification of antigen expression of cell surface antig

ISSN:0248-4900    

DOI:10.1016/0248-4900(93)90117-W

出版商:Blackwell Publishing Ltd    

年代:1993

数据来源: WILEY

摘要:

Summary—A 3‐year interlaboratory proficiency testing for lymphocyte subset phenotyping was initiated as part of the Etalonorme national quality control program. Specimens consisted of fresh whole blood and of lyophilised mononuclear cells (Cytotrol Coulter). The number of participating laboratories was 62 in 1990, 99 in 1991 and 129 in 1992. Statistical analysis indicated that results of phenotyping, expressed as percentages of positive cells, are not related to reagents, instruments or differences in methodology (like, eg time and temperature of incubation). The highest dispersion was observed for total lymphocyte counts and was found to correlate with the type of calibration of the instrument. The coefficients of variation for different lymphocyte subsets were similar if phenotyping was performed on whole blood or lyophilised cells and varied inversely with the percentage of positive cells in each specimen. The consistency of the results indicated that they could serve as a basis for clinical decisi

ISSN:0248-4900    

DOI:10.1016/0248-4900(93)90118-X

出版商:Blackwell Publishing Ltd    

年代:1993

数据来源: WILEY