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The influence of fixation of biological samples on cell count and marker expression stability in flow cytometric analyses
The most common applications of flow cytometry (FC) include diagnostics of haemato-oncological disorders, based on analysis of bone marrow, peripheral blood (PB), or cerebrospinal fluid (CSF) samples. A proper diagnostic process requires standardisation in setting the optimal time frame between mate...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Termedia Publishing House
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7792444/ https://www.ncbi.nlm.nih.gov/pubmed/33456333 http://dx.doi.org/10.5114/ceji.2020.95858 |
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author | SĘDEK, ŁUKASZ KULIS, JAN SŁOTA, ŁUKASZ TWARDOCH, MAGDALENA PIERZYNA-ŚWITAŁA, MAGDALENA PERKOWSKI, BARTOSZ SZCZEPAŃSKI, TOMASZ |
author_facet | SĘDEK, ŁUKASZ KULIS, JAN SŁOTA, ŁUKASZ TWARDOCH, MAGDALENA PIERZYNA-ŚWITAŁA, MAGDALENA PERKOWSKI, BARTOSZ SZCZEPAŃSKI, TOMASZ |
author_sort | SĘDEK, ŁUKASZ |
collection | PubMed |
description | The most common applications of flow cytometry (FC) include diagnostics of haemato-oncological disorders, based on analysis of bone marrow, peripheral blood (PB), or cerebrospinal fluid (CSF) samples. A proper diagnostic process requires standardisation in setting the optimal time frame between material collection and the assay. Unfortunately, this might be difficult to achieve in daily practice due to unintended shipment delays, which might compromise large-scale multicentre studies. Thus, material fixation should be considered as a solution. The most widely used fixative agents are: paraformaldehyde, TransFix(®), Cyto-Chex(®), and serum-containing media. In this review, we attempted to summarise the literature data on the influence of sample storage under different temperatures and times combined with different fixation conditions on the cell count and marker expression levels. Based on the findings of several extensive studies employing fixed PB samples, it can be concluded that the performance of particular fixative greatly depends on the analysed marker and specific PB cell population expressing a given antigen. Preservation of absolute cell count was usually better in Cyto-Chex(®)-fixed PB samples, whereas TransFix(®) tended to better stabilise marker expression levels. CSF-based studies reveal that both serum-containing media and TransFix(®) can prevent cellular loss and enhance FC-based detection of leptomeningeal localisations of haematological malignancies, the latter being more available and having longer shelf-life. As both cell count and marker expression level are the main determinants of quality of biological samples dedicated to FC analyses, it remains to be addressed by the investigators which is the fixative of choice for their specific research aims. |
format | Online Article Text |
id | pubmed-7792444 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Termedia Publishing House |
record_format | MEDLINE/PubMed |
spelling | pubmed-77924442021-01-15 The influence of fixation of biological samples on cell count and marker expression stability in flow cytometric analyses SĘDEK, ŁUKASZ KULIS, JAN SŁOTA, ŁUKASZ TWARDOCH, MAGDALENA PIERZYNA-ŚWITAŁA, MAGDALENA PERKOWSKI, BARTOSZ SZCZEPAŃSKI, TOMASZ Cent Eur J Immunol Review Paper The most common applications of flow cytometry (FC) include diagnostics of haemato-oncological disorders, based on analysis of bone marrow, peripheral blood (PB), or cerebrospinal fluid (CSF) samples. A proper diagnostic process requires standardisation in setting the optimal time frame between material collection and the assay. Unfortunately, this might be difficult to achieve in daily practice due to unintended shipment delays, which might compromise large-scale multicentre studies. Thus, material fixation should be considered as a solution. The most widely used fixative agents are: paraformaldehyde, TransFix(®), Cyto-Chex(®), and serum-containing media. In this review, we attempted to summarise the literature data on the influence of sample storage under different temperatures and times combined with different fixation conditions on the cell count and marker expression levels. Based on the findings of several extensive studies employing fixed PB samples, it can be concluded that the performance of particular fixative greatly depends on the analysed marker and specific PB cell population expressing a given antigen. Preservation of absolute cell count was usually better in Cyto-Chex(®)-fixed PB samples, whereas TransFix(®) tended to better stabilise marker expression levels. CSF-based studies reveal that both serum-containing media and TransFix(®) can prevent cellular loss and enhance FC-based detection of leptomeningeal localisations of haematological malignancies, the latter being more available and having longer shelf-life. As both cell count and marker expression level are the main determinants of quality of biological samples dedicated to FC analyses, it remains to be addressed by the investigators which is the fixative of choice for their specific research aims. Termedia Publishing House 2020-06-08 2020 /pmc/articles/PMC7792444/ /pubmed/33456333 http://dx.doi.org/10.5114/ceji.2020.95858 Text en Copyright © 2020 Termedia http://creativecommons.org/licenses/by-nc-sa/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0). License (http://creativecommons.org/licenses/by-nc-sa/4.0/) |
spellingShingle | Review Paper SĘDEK, ŁUKASZ KULIS, JAN SŁOTA, ŁUKASZ TWARDOCH, MAGDALENA PIERZYNA-ŚWITAŁA, MAGDALENA PERKOWSKI, BARTOSZ SZCZEPAŃSKI, TOMASZ The influence of fixation of biological samples on cell count and marker expression stability in flow cytometric analyses |
title | The influence of fixation of biological samples on cell count and marker expression stability in flow cytometric analyses |
title_full | The influence of fixation of biological samples on cell count and marker expression stability in flow cytometric analyses |
title_fullStr | The influence of fixation of biological samples on cell count and marker expression stability in flow cytometric analyses |
title_full_unstemmed | The influence of fixation of biological samples on cell count and marker expression stability in flow cytometric analyses |
title_short | The influence of fixation of biological samples on cell count and marker expression stability in flow cytometric analyses |
title_sort | influence of fixation of biological samples on cell count and marker expression stability in flow cytometric analyses |
topic | Review Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7792444/ https://www.ncbi.nlm.nih.gov/pubmed/33456333 http://dx.doi.org/10.5114/ceji.2020.95858 |
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