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Updated: Oct 19, 2025
In Situ Exploration of Murine Megakaryopoiesis using Transmission Electron Microscopy
Published on: September 8, 2021
In Situ Exploration of Murine Megakaryopoiesis using Transmission Electron Microscopy
Cyril Scandola1, François Lanza1, Christian Gachet1
1Université de Strasbourg, INSERM, EFS Grand Est, BPPS UMR-S 1255, FMTS.
Abstract:
Differentiation and maturation of megakaryocytes occur in close association with the cellular and extracellular components of the bone marrow. These processes are characterized by the gradual appearance of essential structures in the megakaryocyte cytoplasm such as a polyploid and polylobulated nucleus, an internal membrane network called demarcation membrane system (DMS) and the dense and alpha granules that will be found in circulating platelets. In this article, we describe a standardized protocol for the in situ ultrastructural study of murine megakaryocytes using transmission electron microscopy (TEM), allowing for the identification of key characteristics defining their maturation stage and cellular density in the bone marrow. Bone marrows are flushed, fixed, dehydrated in ethanol, embedded in plastic resin, and mounted for generating cross-sections. Semi-thin and thin sections are prepared for histological and TEM observations, respectively. This method can be used for any bone marrow cell, in any EM facility and has the advantage of using small sample sizes allowing for the combination of several imaging approaches on the same mouse.
Insights
This study presents a standardized transmission electron microscopy (TEM) protocol for examining murine megakaryocytes in bone marrow. The method allows detailed ultrastructural analysis of megakaryocyte maturation and cellular density.
Area of Science:
- Hematology
- Cell Biology
- Microscopy
Background:
- Megakaryocyte differentiation and maturation occur within the bone marrow microenvironment.
- Key maturation events include nuclear changes, development of the demarcation membrane system (DMS), and granule formation.
Purpose of the Study:
- To describe a standardized protocol for in situ ultrastructural study of murine megakaryocytes.
- To enable identification of megakaryocyte maturation stages and cellular density using transmission electron microscopy (TEM).
Main Methods:
- Bone marrow flushing, fixation, ethanol dehydration, and plastic resin embedding.
- Preparation of semi-thin and thin sections for histological and TEM observations.
- Standardized protocol applicable to any bone marrow cell and EM facility.
Main Results:
- Detailed ultrastructural visualization of megakaryocyte maturation.
- Identification of nuclear morphology, demarcation membrane system (DMS), and granule content.
- Assessment of cellular density within the bone marrow.
Conclusions:
- The described TEM protocol provides a standardized method for studying megakaryocyte ultrastructure.
- This technique facilitates the assessment of megakaryocyte maturation and bone marrow cellularity.
- The protocol is versatile, applicable to various bone marrow cells and allows for multimodal imaging from small sample sizes.
