Utilization of imaging flow cytometry to define intermediates of megakaryopoiesis in vivo and in vitro

Journal of Immunological Methods
Kathleen E McGrath

Abstract

Imaging flow cytometry is a particularly powerful analytical approach for the study of megakaryopoiesis. It can utilize well-defined immunophenotypic markers as well as assess maturation of megakaryocytes by their increasing ploidy as they endoreplicate. Imaging flow cytometry can also assess morphometric cell characteristics of size and nuclear to cytoplasmic ratio, which are informative indications of maturation. However, megakaryopoiesis is challenging for flow cytometric analysis, particularly in vivo, because megakaryocytes are very rare in the bone marrow and their odd shape, high DNA content and cell size are similar to clumps of cells. Additionally, both megakaryocytes and immunophenotypically similar platelets are frequently found associated with other cells. Due to these challenges, imaging flow cytometry of megakaryopoiesis exemplifies several strengths of this approach in utilizing fluorescent signal's shape, texture and overlap with other fluorescent signals to distinguish megakaryocytes from a variety of contaminants and to restrict analysis to megakaryocytes, even when associated with other cells. Presented here is a strategy for imaging flow cytometric analysis of rare murine megakaryocytes directly from the bon...Continue Reading

References

Jun 10, 2008·Journal of Immunological Methods·Kathleen E McGrathJames Palis
Feb 5, 2011·Experimental Hematology·Scott A PeslakJames Palis
Jul 5, 2012·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Andrew Filby, Derek Davies
Mar 4, 2014·Blood·Craig N MorrellKristina L Modjeski
Mar 13, 2014·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Lisa M NiswanderJames Palis

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Citations

Oct 24, 2018·Research and Practice in Thrombosis and Haemostasis·Emily C ReddyMargaret L Rand

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