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Akım Sitometride Temel İşlemler ve Tanısal Kullanımı

Year 2025, Volume: 13 Issue: 1, 20 - 26, 28.03.2025

Abstract

Akım sitometrik analiz başta hematolojik malignansiler olmak üzere immün sistem hastalıklarında sıklıkla başvurulan laboratuvar tanı yöntemidir. Gelişen teknolojiye bağlı olarak klinik tanı ve araştırmada uygulama alanı hızla genişlemektedir. Akım sitometrinin öne çıkan başlıca özelliği binlerce hücrenin eş zamanlı, çok parametreli ölçüm ve karakterizasyonunun hızlı bir şekilde yapılabilmesidir. Akım sitometri ile başta kan ve kemik iliği olmak üzere bütün vücut sıvıları, kültür sıvıları, hatta doku örneklerinden elde edilen sıvılarda bulunan hücrelerin analizi mümkündür. Hücresel yapıların canlılık ve gelişim aşamalarının akım sitometri ile izlenmesi mümkündür. Ayrıca, çeşitli mitojenlerle hücresel uyarım sonrasında hücre yüzeylerdeki reseptör ekspresyonları ve fonksiyonlardaki değişimin izlenmesi de mümkündür. Akım sitometrik analizde ilk aşamada FS, SS ve CD45 kapısında hücre farklı hücre gruplarının dağılımı incelenir. Hasta örneklerinin yanısıra sağlıklı kişilerden kan ve kemik iliği örneklerinin akım sitometride incelenmesi patolojik durumların ayırt edilmesinde önemli göstergedir. Teknolojik gelişmelere bağlı olarak akım sitometri yönteminin klinik laboratuvar ve araştırma uygulamalarında kullanımında sık güncellenmeye ihtiyaç duyulmaktadır.

References

  • 1. Rees P, Summers HD, Filby A, Carpenter AE, Doan M. Imaging flow cytometry: a primer. Nat Rev Methods Primers. 2022;2:86.
  • 2. Iyer A, Hamers AAJ, Pillai AB. CyTOF® for the Masses. Front Immunol. 2022;13:815828.
  • 3. Rees P, Summers HD, Filby A, Carpenter AE, Doan M. Imaging flow cytometry: a primer. Nat Rev Methods Primers. 2022;2:86. 4. Gaye Erten, Gülderen Y Demirel, Günnur deniz. Akan hücre ölçer ve uygulama alanları. Medya Tower Yayıncılık Hiz. 2009
  • 5. Abbas AK, Lichtman AH, Pillai S. Cellular and Molecular Immunology. 2021.
  • 6. Rieger AM. Flow cytometry and cell cycle analysis: an overview. Metods Mol Biol. 2022:2579:47-57.
  • 7. Basu K, Green EM, Cheng Y, Craik CS. Why recombinant antibodies - benefits and applications. Curr Opin Biotechnol. 2019;60:153-8.
  • 8. Nordmann GC, Malkemper EP, Landler L, et al. A high sensitivity ZENK monoclonal antibody to map neuronal activity in Aves. Sci Rep 2020;10:915.
  • 9. Ascoli CA, Aggeler B. Overlooked benefits of using polyclonal antibodies. Biotechniques. 2018;65(3):127-36.
  • 10. Kumar R, Parray HA, Shrivastava T, Sinha S, Luthra K. Phage display antibody libraries: A robust approach for generation of recombinant human monoclonal antibodies. Int J Biol Macromol. 2019;135:907-18.
  • 11. Manhas KR, Blattman JN. Flow Cytometry Analysis of Immune Cell Responses. Methods Mol Biol. 2023:2597:105-20.
  • 12. Hays A. Approaches to overcome the challenge of sample stability for flow cytometry analysis in clinical trials.Bioanalysis. 2021;13(21):1587-9.
  • 13. Nolan, JP. Spektral akış sitometrisinin evrimi. Sitometri A. 2022;101:812–7.
  • 14. McKenna RW, Washington LT, Aquino DB, Picker LJ, Kroft SH. Immunophenotypic analysis of hematogones (B-lymphocyte precursors) in 662 consecutive bone marrow specimens by 4-color flow cytometry. Blood. 2001;98(8):2498-507.
  • 15. Nair A, Manohar SM. A flow cytometric journey into cell cycle analysis. Bioanalysis. 2021;13(21):1627-44.
  • 16. McKenna RW, Asplund SL, Kroft SH. Immunophenotypic analysis of hematogones (B-lymphocyte precursors) and neoplastic lymphoblasts by 4-color flow cytometry. Leuk Lymphoma. 2004;45(2):277-85.
  • 17. Heuser M, Freeman SD, Ossenkoppele GJ, et al. 2021 Update on MRD in acute myeloid leukemia: a consensus document from the European LeukemiaNet MRD Working Party. Blood. 2021;138(26):2753-67.
  • 18. Amulic B, Cazalet C, Hayes GL, Metzler KD, Zychlinsky A. Neutrophil function: from mechanisms to disease. Annu Rev Immunol. 2012;30:459-89.
  • 19. Köker N. Kronik granülomatoz hastalıkta nadph oksidaz aktivitesi ve patogenez ilişkisinin yeni tanısal yöntemlerle araştırılması. Doktora tezi. Erciyes Üniversitesi Sağlık Bilimleri Enstitüsü, Kayseri 2023.
  • 20. Şimşek S. Nötrofil Aktivasyon Bozukluğunda Etkilenen İmmünolojik Yolakların Akım Sitometrik Uyarım Testleri İle Araştırılması. Doktora Tezi, Erciyes Üniversitesi Sağlık Bilimleri Enstitüsü, Kayseri 2023:60-62.
  • 21. Akuthota P, Weller PF. Eosinophils and disease pathogenesis. Semin Hematol. 2012;49(2):113-9.
  • 22. Han X, Jorgensen JL, Brahmandam A, et al. Immunophenotypic study of basophils by multiparameter flow cytometry. Arch Pathol Lab Med. 2008;132(5):813-9.
  • 23. Santos AF, Alpan O, Hoffmann HJ. Basophil activation test: Mechanisms and considerations for use in clinical trials and clinical practice. Allergy. 2021;76(8):2420-32.
  • 24. Eberlein B. Basophil Activation as Marker of Clinically Relevant Allergy and Therapy Outcome. Front Immunol. 2020;11:1815.
  • 25. Auffray C, Sieweke MH, Geissmann F. Blood monocytes: development, heterogeneity, and relationship with dendritic cells. Annu Rev Immunol. 2009;27:669-92.
  • 26. Köker MY. Diagnostic Management of Chronic Granulomatous Disease. Asthma Allergy Immunol 2023;21:1-2.
  • 27. Lanzavecchia A, Sallusto F. Human B cell memory. Curr Opin Immunol. 2009;21(3):298-304.

Fundamental Flow Cytometry Procedures and Application for Diagnosis

Year 2025, Volume: 13 Issue: 1, 20 - 26, 28.03.2025

Abstract

Flow cytometric analysis is a laboratory diagnostic method frequently used in immune system diseases, especially hematological malignancies. Its application area in clinical diagnosis and research is rapidly expanding due to developing technology. The main feature of flow cytometry is that it can rapidly perform simultaneous, multi-parameter measurement and characterization of thousands of cells. With flow cytometry, it is possible to analyze cells in all body fluids, especially blood and bone marrow, culture fluids, and even fluids obtained from tissue samples. It is possible to monitor the vitality and developmental stages of cellular structures with flow cytometry. In addition, it is also possible to monitor the changes in receptor expressions and functions on cell surfaces after cellular stimulation with various mitogens. In flow cytometric analysis, the distribution of different cell groups in the FS, SS and CD45 gates is examined in the first stage. In addition to patient samples, examination of blood and bone marrow samples from healthy individuals by flow cytometry is an important indicator in distinguishing pathological conditions. Depending on technological developments, the use of flow cytometry method in clinical laboratory and research applications requires frequent updates.

References

  • 1. Rees P, Summers HD, Filby A, Carpenter AE, Doan M. Imaging flow cytometry: a primer. Nat Rev Methods Primers. 2022;2:86.
  • 2. Iyer A, Hamers AAJ, Pillai AB. CyTOF® for the Masses. Front Immunol. 2022;13:815828.
  • 3. Rees P, Summers HD, Filby A, Carpenter AE, Doan M. Imaging flow cytometry: a primer. Nat Rev Methods Primers. 2022;2:86. 4. Gaye Erten, Gülderen Y Demirel, Günnur deniz. Akan hücre ölçer ve uygulama alanları. Medya Tower Yayıncılık Hiz. 2009
  • 5. Abbas AK, Lichtman AH, Pillai S. Cellular and Molecular Immunology. 2021.
  • 6. Rieger AM. Flow cytometry and cell cycle analysis: an overview. Metods Mol Biol. 2022:2579:47-57.
  • 7. Basu K, Green EM, Cheng Y, Craik CS. Why recombinant antibodies - benefits and applications. Curr Opin Biotechnol. 2019;60:153-8.
  • 8. Nordmann GC, Malkemper EP, Landler L, et al. A high sensitivity ZENK monoclonal antibody to map neuronal activity in Aves. Sci Rep 2020;10:915.
  • 9. Ascoli CA, Aggeler B. Overlooked benefits of using polyclonal antibodies. Biotechniques. 2018;65(3):127-36.
  • 10. Kumar R, Parray HA, Shrivastava T, Sinha S, Luthra K. Phage display antibody libraries: A robust approach for generation of recombinant human monoclonal antibodies. Int J Biol Macromol. 2019;135:907-18.
  • 11. Manhas KR, Blattman JN. Flow Cytometry Analysis of Immune Cell Responses. Methods Mol Biol. 2023:2597:105-20.
  • 12. Hays A. Approaches to overcome the challenge of sample stability for flow cytometry analysis in clinical trials.Bioanalysis. 2021;13(21):1587-9.
  • 13. Nolan, JP. Spektral akış sitometrisinin evrimi. Sitometri A. 2022;101:812–7.
  • 14. McKenna RW, Washington LT, Aquino DB, Picker LJ, Kroft SH. Immunophenotypic analysis of hematogones (B-lymphocyte precursors) in 662 consecutive bone marrow specimens by 4-color flow cytometry. Blood. 2001;98(8):2498-507.
  • 15. Nair A, Manohar SM. A flow cytometric journey into cell cycle analysis. Bioanalysis. 2021;13(21):1627-44.
  • 16. McKenna RW, Asplund SL, Kroft SH. Immunophenotypic analysis of hematogones (B-lymphocyte precursors) and neoplastic lymphoblasts by 4-color flow cytometry. Leuk Lymphoma. 2004;45(2):277-85.
  • 17. Heuser M, Freeman SD, Ossenkoppele GJ, et al. 2021 Update on MRD in acute myeloid leukemia: a consensus document from the European LeukemiaNet MRD Working Party. Blood. 2021;138(26):2753-67.
  • 18. Amulic B, Cazalet C, Hayes GL, Metzler KD, Zychlinsky A. Neutrophil function: from mechanisms to disease. Annu Rev Immunol. 2012;30:459-89.
  • 19. Köker N. Kronik granülomatoz hastalıkta nadph oksidaz aktivitesi ve patogenez ilişkisinin yeni tanısal yöntemlerle araştırılması. Doktora tezi. Erciyes Üniversitesi Sağlık Bilimleri Enstitüsü, Kayseri 2023.
  • 20. Şimşek S. Nötrofil Aktivasyon Bozukluğunda Etkilenen İmmünolojik Yolakların Akım Sitometrik Uyarım Testleri İle Araştırılması. Doktora Tezi, Erciyes Üniversitesi Sağlık Bilimleri Enstitüsü, Kayseri 2023:60-62.
  • 21. Akuthota P, Weller PF. Eosinophils and disease pathogenesis. Semin Hematol. 2012;49(2):113-9.
  • 22. Han X, Jorgensen JL, Brahmandam A, et al. Immunophenotypic study of basophils by multiparameter flow cytometry. Arch Pathol Lab Med. 2008;132(5):813-9.
  • 23. Santos AF, Alpan O, Hoffmann HJ. Basophil activation test: Mechanisms and considerations for use in clinical trials and clinical practice. Allergy. 2021;76(8):2420-32.
  • 24. Eberlein B. Basophil Activation as Marker of Clinically Relevant Allergy and Therapy Outcome. Front Immunol. 2020;11:1815.
  • 25. Auffray C, Sieweke MH, Geissmann F. Blood monocytes: development, heterogeneity, and relationship with dendritic cells. Annu Rev Immunol. 2009;27:669-92.
  • 26. Köker MY. Diagnostic Management of Chronic Granulomatous Disease. Asthma Allergy Immunol 2023;21:1-2.
  • 27. Lanzavecchia A, Sallusto F. Human B cell memory. Curr Opin Immunol. 2009;21(3):298-304.
There are 26 citations in total.

Details

Primary Language Turkish
Subjects Pediatric Immunology and Allergic Diseases
Journal Section Reviews
Authors

Sevil Şimşek 0000-0001-5998-6917

Nezihe Köker 0000-0002-3962-9397

Huriye Çelikzencir 0009-0006-8538-5676

Mustafa Yavuz Köker 0000-0001-7061-8525

Publication Date March 28, 2025
Submission Date December 1, 2024
Acceptance Date March 23, 2025
Published in Issue Year 2025 Volume: 13 Issue: 1

Cite

Vancouver Şimşek S, Köker N, Çelikzencir H, Köker MY. Akım Sitometride Temel İşlemler ve Tanısal Kullanımı. pediatr pract res. 2025;13(1):20-6.