Maria Faresjö , Junko Johansson , Ulrika Islander , Andrea Tompa
{"title":"Flow cytometry reveals constant lymphocyte proportions after long-term cryopreservation of whole blood in TransFix® cell stabilization reagent","authors":"Maria Faresjö , Junko Johansson , Ulrika Islander , Andrea Tompa","doi":"10.1016/j.jim.2025.113853","DOIUrl":null,"url":null,"abstract":"<div><div>Flow cytometry is an important technique for characterization of immune cells, with accurate lymphocyte profiling being essential for clinical diagnostics and research applications. While immediate processing of blood samples is ideal, long-term storage solutions are needed for large-scale studies or settings without immediate access to laboratory facilities. TransFix® is a chemical stabilization solution that allows delayed analysis by preserving cell morphology and surface markers. However, the impact of long-term cryopreservation in TransFix® on lymphocyte integrity remains underexplored. In this study, we evaluated the efficacy of cryopreservation in TransFix® for maintaining the proportions of key lymphocyte subsets, including CD3<sup>+</sup> T cells, CD3<sup>+</sup>CD4<sup>+</sup> T helper cells, CD3<sup>+</sup>CD8<sup>+</sup> cytotoxic T cells, CD19<sup>+</sup> B cells, and CD16<sup>+</sup>/CD56<sup>+</sup> natural killer cells. Blood samples were cryopreserved in TransFix® for varying time periods, up to 48 months, and compared to fresh samples using flow cytometry. The results show that the proportions of lymphocyte subsets remain stable during cryopreservation for up to 48 months, with no significant differences observed between fresh and cryopreserved samples. This suggests that TransFix® can successfully preserve lymphocyte integrity for long-term storage, providing a reliable option for delayed analysis. These results highlight the usefulness of TransFix® for studies that require extended storage, making it easier to conduct immune monitoring in a wide range of settings.</div></div>","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":"539 ","pages":"Article 113853"},"PeriodicalIF":1.6000,"publicationDate":"2025-03-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of immunological methods","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0022175925000535","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0
Abstract
Flow cytometry is an important technique for characterization of immune cells, with accurate lymphocyte profiling being essential for clinical diagnostics and research applications. While immediate processing of blood samples is ideal, long-term storage solutions are needed for large-scale studies or settings without immediate access to laboratory facilities. TransFix® is a chemical stabilization solution that allows delayed analysis by preserving cell morphology and surface markers. However, the impact of long-term cryopreservation in TransFix® on lymphocyte integrity remains underexplored. In this study, we evaluated the efficacy of cryopreservation in TransFix® for maintaining the proportions of key lymphocyte subsets, including CD3+ T cells, CD3+CD4+ T helper cells, CD3+CD8+ cytotoxic T cells, CD19+ B cells, and CD16+/CD56+ natural killer cells. Blood samples were cryopreserved in TransFix® for varying time periods, up to 48 months, and compared to fresh samples using flow cytometry. The results show that the proportions of lymphocyte subsets remain stable during cryopreservation for up to 48 months, with no significant differences observed between fresh and cryopreserved samples. This suggests that TransFix® can successfully preserve lymphocyte integrity for long-term storage, providing a reliable option for delayed analysis. These results highlight the usefulness of TransFix® for studies that require extended storage, making it easier to conduct immune monitoring in a wide range of settings.
期刊介绍:
The Journal of Immunological Methods is devoted to covering techniques for: (1) Quantitating and detecting antibodies and/or antigens. (2) Purifying immunoglobulins, lymphokines and other molecules of the immune system. (3) Isolating antigens and other substances important in immunological processes. (4) Labelling antigens and antibodies. (5) Localizing antigens and/or antibodies in tissues and cells. (6) Detecting, and fractionating immunocompetent cells. (7) Assaying for cellular immunity. (8) Documenting cell-cell interactions. (9) Initiating immunity and unresponsiveness. (10) Transplanting tissues. (11) Studying items closely related to immunity such as complement, reticuloendothelial system and others. (12) Molecular techniques for studying immune cells and their receptors. (13) Imaging of the immune system. (14) Methods for production or their fragments in eukaryotic and prokaryotic cells.
In addition the journal will publish articles on novel methods for analysing the organization, structure and expression of genes for immunologically important molecules such as immunoglobulins, T cell receptors and accessory molecules involved in antigen recognition, processing and presentation. Submitted full length manuscripts should describe new methods of broad applicability to immunology and not simply the application of an established method to a particular substance - although papers describing such applications may be considered for publication as a short Technical Note. Review articles will also be published by the Journal of Immunological Methods. In general these manuscripts are by solicitation however anyone interested in submitting a review can contact the Reviews Editor and provide an outline of the proposed review.