eISSN: 1644-4124
ISSN: 1426-3912
Central European Journal of Immunology
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2/2020
vol. 45
 
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abstract:
Review paper

The influence of fixation of biological samples on cell count and marker expression stability in flow cytometric analyses

Łukasz Sędek
1
,
Jan Kulis
2
,
Łukasz Słota
2
,
Magdalena Twardoch
2
,
Magdalena Pierzyna-Świtała
2
,
Bartosz Perkowski
2
,
Tomasz Szczepański
2

  1. Department of Microbiology and Immunology, School of Medicine with the Division of Dentistry in Zabrze, Medical University of Silesia in Katowice, Zabrze, Poland
  2. Department of Paediatric Haematology and Oncology, School of Medicine with the Division of Dentistry in Zabrze, Medical University of Silesia in Katowice, Zabrze, Poland
Cent Eur J Immunol 2020; 45 (2): 206-213
DOI: https://doi.org/10.5114/ceji.2020.95858
Online publish date: 2020/06/08
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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.
keywords:

flow cytometry, peripheral blood, cerebrospinal fluid, fixative, Transfix, Cyto-Chex, antigen expression, cryopreservation
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