Analysis of Erythropoiesis Using Imaging Flow Cytometry

Methods in Molecular Biology
Theodosia A Kalfa, Kathleen E McGrath

Abstract

Erythroid maturation has been classically defined based on the remarkable changes visualized through microscopy. These involve the decrease in cell size, nuclear condensation and organelle loss, and include the final unique asymmetric division creating the short-lived nucleated pyrenocyte and the enucleate reticulocyte that matures into the red blood cell. Understanding the regulation of these processes has been challenging due to the difficulty in obtaining sufficient numbers of cells, particularly of rare intermediates, to study by microscopy. While flow cytometry can provide quantitative analysis of high cell numbers as well as critical tools for assaying processes like cell cycle, apoptosis and cell signaling, it cannot analyze or categorize cells based on morphology. Imaging flow cytometry (IFC) combines microscopy and flow cytometry by capturing brightfield and fluorescent images of large numbers of cells, which can be quantitated for both morphometric and fluorescent characteristics. Over the past 10 years, this approach has been increasingly used to study aspects of erythropoiesis. This chapter describes how to utilize IFC to enumerate multiple specific stages of erythropoiesis from primary tissue, as well as how to cul...Continue Reading

Citations

Nov 20, 2020·Blood Advances·Hakim Ouled-HaddouLoïc Garçon
Nov 14, 2020·Science·Junyue CaoJay Shendure
May 8, 2021·Stem Cell Research & Therapy·Tso-Fu WangDer-Shan Sun
Aug 29, 2020·Haematologica·Sara El HossWassim El Nemer
Dec 19, 2021·Nature Communications·Daniel HidalgoMerav Socolovsky

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