A Comparative Study Of Bone Marrow Aspiration Smear And Cell Block Using Romanwasky And Hematoxylin And Eosin Stain
U Uniya, K Likhar, R Nigam
bone marrow, cell blocks, romanowsky stain.
U Uniya, K Likhar, R Nigam. A Comparative Study Of Bone Marrow Aspiration Smear And Cell Block Using Romanwasky And Hematoxylin And Eosin Stain. The Internet Journal of Pathology. 2012 Volume 13 Number 3.
The bone marrow is one of the body’s largest organs. It constitutes 4.5% of the total body weight and weights 3375 grams in a 75 kg individual1. The hematopoietic bone marrow is organized around the vasculature of the bone cavity. The main function of the bone marrow is to supply mature hematopoietic cells for circulating blood in a steady-state as well as to respond to increased physiologic or pathologic demands.
Recent advances in the treatment of hematologic malignancies have been paralleled by renewed interest on the part of pathologists and hematologists in methods of obtaining and preparing bone marrow for diagnostic studies.
Bone marrow aspirate is a cytologic preparation of bone marrow cells obtained by aspiration of marrow and a smear of the cells. Bone marrow aspirate smears are primarily used for the assessment of differentia count, maturational status, morphologic details, myeloid-to-erythroid (M:E) ratio. Bone marrow aspirated cell blocks provide a better and more detailed estimate of bone marrow cellularity. They are particularly useful for patients with aplastic / hypoplastic marrow. Aspiration of the marrow has been utilized from cytologic assessment, with analysis directed toward morphology and obtainment of differential cell count.
In H&E stained sections of bone marrow, the more mature stages of the erythroid and myeloid cells, adipocytes and megakaryocytes can be identified. However lymphoid cells as well as immature progenitor cells cannot be reliably identified. The use of H&E and Romanowsky group of stains in the bone marrow analysis are very important and allow for studies of marrow’s overall cellularity, detection of focal lesions and extent of infiltration by various pathological entities.
Bone marrow studies aid in the diagnosis, staging and monitoring of several diseases Thus the present study was being conducted to compare efficacy & to explore further utility of Geimsa and Leishman stains of Romanowsky group in bone marrow aspiration smears.
The present study was undertaken with the objectives –
Materials And Methods
The present study was done in Hamidia Hospital, Bhopal. Department of pathology from May 2008 to Sep. 2009. A consecutive marrow samples from 83 patients attending hematology OPD or ward of Hamidia Hospital, Bhopal. In the present study included 83 cases in which complete peripheral smears, bone marrow and cell blocks was available. The selection of cases was based on the clinical examination and peripheral blood smear of patients in which a hematological disorder was suspected.
History and clinical examination was done as per proforma. Investigations like Hemoglobin, Total WBC count, Platelet count, Reticulocyte count, Peripheral blood smear examination, Blood glucose, Blood urea, Serum creatinine, AST, ALT, Bone marrow aspiration were performed
Bone Marrow Aspiration was done through safe and preferred sites. The preffered site for young and old patients
Other sites for Bone Marrow Aspiration4 are Posterior superior iliac spine, Posterior iliac crest, Anterior iliac crest, Spinous process of lumber vertebra, Upper end of tibia. The sternum remains the most commonly employed aspiration site.
Cell blocks preparation
The clot section is prepared from the blood left over after aspirate smears have been made. The blood containing admixed marrow particles is transferred to a container with 1ml of 10% formal saline and 2ml methanol for fixation. Future processing is carried out as for routine histopathological staining.
The slides were viewed and reported. The reporting was done in conventional way as well as a scoring system created indigenously
The maximum score for a single case, taking into account all the four parameters was 12. Thus the maximum possible score in the study was calculated by multiplying the number of cases by 12 for each of 2 stains. A “quality index” was obtained by finding out the ratio of actual score obtained to the maximum score possible.
The quality index for each of two stains was compared. As the smears of all the cases were stained with Leishman stain and Geimsa stain, Quality Index of all slides were calculated and compared.
The present study was conducted for comparison between bone marrow aspirate smears stained by Romanowsky group of stains (Leishman & Giemsa) and paraffin embedded cell block section stained by Hematoxylin & Eosin stain. Total 83 cases were selected in which complete peripheral smears, bone marrow aspirate and cell blocks were available. The findings obtained are as follows-
By Romanowsky group of stains and Hematoxylin & Eosin stained cellblocks, diagnosis of various hematological disorders was made. Out of 83 cases studied there were 60 cases (72.2%) of anaemia, 9 (10.8%) platelet disorders, 6 (7.2%) Acute Leukaemias, 3(3.6%) shift to left, 2 (2.4%) Lymph proliferative disorders, 1 (1.2%) MDS, 1(1.2%) Leishmaniasis and 1 (1.2%) hyperspleenism. Maximum cases 40 were in age group of 11-50 years of age in megaloblastic anaemia. 32 (69.5%) cases in megaloblastic anaemia showing male preponderance.
In Leishman and Giemsa stained smears parameters studied as per background of smears, overall staining, cell morphology and nuclear details, showed that except nuclear details all parameters are alike. In nuclear details the Quality was superior in Leishman than Giemsa. Quality Index in Leishman stain was 0.20 and in Giemsa stain was 0.12.
The cellularities analyzed on bone marrow aspiration showed maximum 84.3% were hypercellular marrow. Cell blocks help in better appreciation of hypocellular / acellular marrow. Megakaryocytic & Eosinophilic precursors are better identifiable in H &E stained block.
The purpose of this work is to compare efficacy of Romanowsky group of stains (Geimsa and Leishman ) in bone marrow aspiration smears & to assess the role of paraffin embedded cell block of bone marrow aspirates in bone marrow examination. In nuclear details the Quality was superior in Leishman than Giemsa. Quality Index in Leishman stain was 0.20 and in Giemsa stain was 0.12.
The cases diagnosed as megaloblastic anemia showed that male female ratio was 2:1 and the commonest age group was 21 to 30 years. This is in variance with studies of
The overall staining quality is superior in Leishman stain than in Giemsa stain.Hematoxylin & Eosin stained cell blocks are particularly helpful in hypo cellular marrow aspirate. Megakaryocytes & eosinophillic precursors can be better appreciated in cell block sections. Cellblocks can be stored and used for specialized procedure like immunohistochemisty in future.