**Cytopathology of MDS/MPN and AML by H&E Staining**

**Cytopathology of MDS/MPN and AML by H&E Staining**

DOI: 10.5772/intechopen.71567

#### Tatsuaki Tsuruyama Tatsuaki Tsuruyama Additional information is available at the end of the chapter

Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/intechopen.71567

#### **Abstract**

Bone marrow (BM) clots are routinely sampled in aspiration tests, and their sections are prepared for histological observation by hematoxylin and eosin (H&E) staining. However, H&E-stained sections are considered less informative than those stained by the May-Grünwald Giemsa (M-G) stain; thus, diagnosis using H&E-stained clot samples is challenging for pathologists. In fact, the diagnostic evaluation is limited to the observation of cellular morphology and the myeloid-erythroid cell ratio. Pathologists leave cellular observation to laboratory hematologists, who generally use M-G staining. In this chapter, the utility of bone marrow clot specimens for diagnosis by H&E staining is reviewed. Specifically, the review provides a descriptive and illustrative explanation of the diagnosis of acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), and myelodysplastic syndrome/myelocytic proliferative neoplasm (MDS/MPN) and demonstrates the possibility of diagnosis on the basis of the characteristic features of blast cells. Clot specimens appear to be useful for the diagnosis of hematopoietic dysplasia by pathologists, and this approach can provide more informative findings for hematologists.

**Keywords:** bone marrow clots, hematoxylin and eosin, acute myeloid leukemia

#### **1. Introduction**

Bone marrow (BM) clots and their specimens can be evaluated histopathologically by hematoxylin and eosin (H&E) staining [1, 2]. However, this method is considered to yield limited information regarding cellular morphology, myeloid-erythroid cell ratio, size and morphology of megakaryocytes [3], increase in multiple myeloma [4], involvement of lymphoma, and metastatic tumor such as neuroblastoma [5–7]. In particular, BM clot sections comprising sinusoidal blood are inadequate for morphological interpretation [2]. However, the cellblock technique ensures the capture of cells required for diagnosis, and laboratories routinely apply this sample preparation technique for enhancing the usability of the BM clot specimen [2, 8].

Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. © 2018 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

© 2016 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons

To assist in morphologic diagnosis, immunohistochemistry (IHC) is available for assessment and immunophenotyping. For instance, by IHC, the marker glycophorin can assess the myeloid-erythroid cell ratio, and spectrin can efficiently detect erythroid cellular lineage [9]. P53 can also be used for differential diagnosis between refractory anemia and aplastic anemia [10]. An international Working Party for the Standardization of Bone Marrow IHC was formed by the International Council for Standardization in Hematology to prepare a set of guidelines for the standardization of handling BM specimens and reports [11]. While the histological examination of BM introduces new aspects of blood disease, the diagnosis of hematopoietic disorders using H&E-stained BM clot sections remains a challenge for nonhematopathologists. In fact, hematopoietic disorders have been diagnosed using May-Grünwald Giemsa (M-G) staining by hematologists and pathologists, and the findings obtained from H&E-stained BM clot sections are used as a reference for those obtained from M-G–stained samples.

This review discusses the information obtained from H&E-stained BM clot sections and its advantages and limitations over M-G–stained BM clot sections. The techniques are discussed with respect to examination of erythrocytes and myelocytes and the diagnosis of acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), and myelodysplastic syndrome/myelocytic proliferative neoplasm (MDS/MPN).

**Figure 1.** Morphology of erythroblasts and myeloblasts. (A) an erythroblast showing a transparent nucleus with condensed nucleoli (600×). (B) a myeloblast showing a transparent nucleus with one prominent eosinophilic nucleolus. The cytoplasm is eosinophilic, rough, and heterogeneously condensed (600×). (C) Two megakaryoblasts show transparent and lobulated nuclei with prominent eosinophilic nucleoli. The cytoplasm is eosinophilic, rough, and heterogeneously

Cytopathology of MDS/MPN and AML by H&E Staining http://dx.doi.org/10.5772/intechopen.71567 11

**Figure 2.** Morphology of myelocyte lineages detected by H&E staining. Immature neutrophil lineages (A–C) and immature eosinophil lineages (D). (A) A pro-myelocyte showing a nucleus surrounded by halo (represented by an arrow). Nucleoli are not prominent relative to myeloblasts. The cytoplasm is eosinophilic, rough, and heterogeneously condensed (600×). (B) A myelocyte showing a regular oval nucleus with a contour and without indentation. The cytoplasm is eosinophilic, rough, and heterogeneously condensed (600×). (C) A meta-myelocyte showing a U-shaped or bent nucleus. The cytoplasm is eosinophilic, rough, and heterogeneously condensed. No nucleoli are visible (600×). (D) Variable immature eosinophil lineages. The arrow indicates a pro-myelocyte, and the arrowhead indicates a myelocyte.

condensed. The edges are irregular, with a number of villi (600×).

A mature eosinophil can be seen in the upper-left corner (600×).
