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Updated: May 2, 2026
Database-guided Flow-cytometry for Evaluation of Bone Marrow Myeloid Cell Maturation
Published on: November 3, 2018
Improved quantitative analysis of primary bone marrow megakaryocytes utilizing imaging flow cytometry
Lisa M Niswander1, Kathleen E McGrath, John C Kennedy
1Department of Pediatrics, Center for Pediatric Biomedical Research, University of Rochester Medical Center, Rochester, New York, 14642; Department of Pathology and Laboratory Medicine, University of Rochester Medical Center, Rochester, New York, 14642.
Abstract:
Life-threatening thrombocytopenia can develop following bone marrow injury due to decreased platelet production from megakaryocytes (MKs). However, the study of primary MKs has been complicated by their low frequency in the bone marrow and by technical challenges presented by their unique maturation properties. More accurate and efficient methods for the analysis of in vivo MKs are needed to enhance our understanding of megakaryopoiesis and ultimately develop new therapeutic strategies for thrombocytopenia. Imaging flow cytometry (IFC) combines the morphometric capabilities of microscopy with the high-throughput analyses of flow cytometry (FC). Here, we investigate the application of IFC on the ImageStream(X) platform to the analysis of primary MKs isolated from murine bone marrow. Our data highlight and address technical challenges for conventional FC posed by the wide range of cellular size within the MK lineage as well as the shared surface phenotype with abundant platelet progeny. We further demonstrate that IFC can be used to reproducibly and efficiently quantify the frequency of primary murine MKs in the marrow, both at steady-state and in the setting of radiation-induced bone marrow injury, as well as assess their ploidy distribution. The ability to accurately analyze the full spectrum of maturing MKs in the bone marrow now allows for many possible applications of IFC to enhance our understanding of megakaryopoiesis and platelet production.
Insights
Imaging flow cytometry (IFC) offers a new method to study megakaryocytes (MKs), crucial for platelet production. This technique overcomes challenges in analyzing MKs, aiding research into thrombocytopenia.
Area of Science:
- Hematology
- Cell Biology
- Biotechnology
Background:
- Thrombocytopenia, a condition of low platelet count, often results from bone marrow injury and impaired megakaryocyte (MK) function.
- Studying primary MKs is difficult due to their rarity in bone marrow and complex maturation, hindering the development of effective thrombocytopenia treatments.
Purpose of the Study:
- To evaluate the utility of imaging flow cytometry (IFC) for analyzing primary murine megakaryocytes (MKs).
- To address existing technical limitations in conventional flow cytometry for MK analysis.
- To establish a reproducible method for quantifying MKs and their ploidy in vivo.
Main Methods:
- Isolation of primary MKs from murine bone marrow.
- Application of imaging flow cytometry (IFC) on the ImageStream(X) platform.
- Analysis of MKs at steady-state and following radiation-induced bone marrow injury.
Main Results:
- IFC effectively addresses challenges posed by MK size variation and shared surface markers with platelets.
- IFC enables reproducible and efficient quantification of primary murine MK frequency.
- IFC accurately assesses MK ploidy distribution in both steady-state and injury conditions.
Conclusions:
- IFC provides a powerful tool for analyzing the full spectrum of maturing MKs in the bone marrow.
- This method enhances the understanding of megakaryopoiesis and platelet production.
- IFC applications hold promise for developing novel therapeutic strategies for thrombocytopenia.
