Abstract
This chapter covers the principle of flow cytometry (FCM), the techniques and clinical applications of it. FCM provides us very quick assessment of cell surface antigens, DNA content and intracellular proteins. The instrument is now used not only for research but also for routine clinical activities. Both histology and cytology samples can be processed for FCM. However cytology samples are easy to process due to less effort for disaggregation. The chapter describes the positive and negative control of FCM, steps of sample preparation, acquisition and data interpretation. The sub-classification of lymphomas and other diagnostic use of FCM in exfoliative cytology also have been discussed. FCM is one of the important technologies to detect the apoptotic cell death. Quantitation of the apoptotic cell is possible with the help of light scatter (a loss in forward light scatter), plasma membrane changes and DNA content (a sub-diploid peak). The steps of FCM for the detection of apoptosis are also described here.
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References
Saikia UN, Dey P, Vohra H, Gupta SK. DNA flow cytometry of non Hodgkin’s lymphomas: correlation with cytologic grade and clinical relapse. Diagn Cytopathol. 2000;22:153–6.
Kentrou NA, Tsagarakis NJ, Tzanetou K, Damala M, Papadimitriou KA, Skoumi D, Stratigaki A, Anagnostopoulos NI, Malamou-Lada E, Athanassiadou P, Paterakis G. An improved flow cytometric assay for detection and discrimination between malignant cells and atypical mesothelial cells, in serous cavity effusions. Cytometry B Clin Cytom. 2011;80(5):324–34.
Dey P, Amir T, Al Jassar A, et al. Combined applications of fine needle aspiration cytology and flow cytometric immunphenotyping for diagnosis and classification of non Hodgkin lymphoma. Cytojournal. 2006;3:24.
Johansson U, Bloxham D, Couzens S, Jesson J, Morilla R, Erber W, Macey M, British Committee for Standards in Haematology. Guidelines on the use of multicolour flow cytometry in the diagnosis of haematological neoplasms. British Committee for Standards in Haematology. Br J Haematol. 2014;165(4):455–88.
Chang Q, Hedley D. Emerging applications of flow cytometry in solid tumor biology. Methods. 2012;57(3):359–67.
Adan A, Alizada G, Kiraz Y, Baran Y, Nalbant A. Flow cytometry: basic principles and applications. Crit Rev Biotechnol. 2017;37(2):163–76.
Diaz-Arias AA, Loy TS, Bickel JT, et al. Utility of Ber-EP4 in the diagnosis of adenocarcinoma in effusions:an immunocytochemical study of 232 cases. Diagn Cytopathol. 1993;9:516–21.
Sánchez-Carbayo M, Ciudad J, Urrutia M, Navajo JA, Orfao A. Diagnostic performance of the urinary bladder carcinoma antigen ELISA test and multiparametric DNA/cytokeratin flow cytometry in urine voided samples from patients with bladder carcinoma. Cancer. 2001;92(11):2811–9.
Yu GH, Vergara N, Moore EM, King RL. Use of flow cytometry in the diagnosis of lymphoproliferative disorders in fluid specimens. Diagn Cytopathol. 2014;42(8):664–70.
Bromberg JE, Breems DA, Kraan J, Bikker G, van der Holt B, Smitt PS, van den Bent MJ, van’t Veer M, Gratama JW. CSF flow cytometry greatly improves diagnostic accuracy in CNS hematologic malignancies. Neurology. 2007;68(20):1674–9.
Coon JS, Landay AL, Weinstein RS. Advances in flow cytometry for diagnostic pathology. Lab Investig. 1987;57:453–79.
Zola H, Swart B, Nicholson I, et al. CD molecules 2005: human cell differentiation molecules. Blood. 2005;106:3123–6.
Dey P. The role of ancillary techniques to diagnose and sub-classifiy non Hodgkin lymphomas on fine needle aspiration cytology. Cytopathology. 2006;17(5):275–87.
Darzynkiewicz Z, Bedner E, Smolewski P. Flow cytometry in analysis of cell cycle and apoptosis. Semin Hematol. 2001;38(2):179–93.
Darzynkiewicz Z, Bruno S, Del Bino G, Gorczyca W, Hotz MA, Lassota P, Traganos F. Features of apoptotic cells measured by flow cytometry. Cytometry. 1992;13(8):795–808.
Riley RS, Ben-Ezra JM, Tidwell A, Romagnoli G. Reticulocyte analysis by flow cytometry and other techniques. Hematol Oncol Clin North Am. 2002;16(2):373–420.
Miyoshi H, Arakawa F, Sato K, Kimura Y, Kiyasu J, Takeuchi M, Yoshida M, Ichikawa A, Ishibashi Y, Nakamura Y, Nakashima S, Niino D, Sugita Y, Ohshima K. Comparison of CD20 expression in B-cell lymphoma between newly diagnosed, untreated cases and those after rituximab treatment. Cancer Sci. 2012;103(8):1567–73.
Chatterjee T, Mallhi RS, Venkatesan S. Minimal residual disease detection using flow cytometry: applications in acute leukemia. Med J Armed Forces India. 2016;72(2):152–6.
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Dey, P. (2018). Flow Cytometry: Basic Principles, Procedure and Applications in Pathology. In: Basic and Advanced Laboratory Techniques in Histopathology and Cytology. Springer, Singapore. https://doi.org/10.1007/978-981-10-8252-8_17
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DOI: https://doi.org/10.1007/978-981-10-8252-8_17
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