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A Comparative Study of Bone Marrow Aspiration Smear and Cell Block Using Romanwasky and Hematoxylin and Eosin Stain U Uniya, K Likhar, R Nigam

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A Comparative Study of Bone Marrow Aspiration Smear and Cell Block Using Romanwasky and Hematoxylin and Eosin Stain U Uniya, K Likhar, R Nigam The Internet Journal of Pathology ISPUB.COM Volume 13 Number 3 A Comparative Study Of Bone Marrow Aspiration Smear And Cell Block Using Romanwasky And Hematoxylin And Eosin Stain U Uniya, K Likhar, R Nigam Citation 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. Abstract Background: Bone marrow aspirate is a cytologic preparation of bone marrow cells obtained by aspiration of marrow and a smear of the cells. Aspiration of the marrow has been utilized for cytological assessment, with analysis directed toward morphology and differential cell count. 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. Objectives: To compare efficacy of Romanowsky group of stains (Geimsa and Leishman) in bone marrow aspiration smears with paraffin embedded cell blocks section. Methods: The present study was done Hamidia Hospital, Bhopal. Department of Pathology from May 2008 to Sep. 2009. 83 cases in which complete peripheral blood smears, bone marrow and cell blocks was available were included in the study. Bone marrow aspiration was performed, Cell block was prepared & smears were stained with Leishman stain and Geimsa stain. Quality Index of all slides were calculated and compared. Results: Comparison was done between bone marrow aspirate smears stained by Romanowsky group of stains (Leishman & Giemsa) and paraffin embedded cell block section stained by Hematoxylin & Eosin stain. Out of 83 cases, there were 60 cases (72.2%) of anaemias, 9 (10.8%) platelet disorders, 6 (7.2%) Acute Leukaemias, 3(3.6%) shift to left, 2 (2.4%) Lymphoproliferative disorders, 1 (1.2%) MDS, 1(1.2%) Leishmaniasis and 1 (1.2%) hyperspleenism. Conclusions: The overall staining quality is superior in Leishman stain than in Giemsa stain. H&E stained are particularly helpful in hypocellular marrow aspirate. INTRODUCTION for the assessment of differentia count, maturational status, The bone marrow is one of the body’s largest organs. It morphologic details, myeloid-to-erythroid (M:E) ratio. Bone constitutes 4.5% of the total body weight and weights 3375 marrow aspirated cell blocks provide a better and more grams in a 75 kg individual1. The hematopoietic bone detailed estimate of bone marrow cellularity. They are marrow is organized around the vasculature of the bone particularly useful for patients with aplastic / hypoplastic cavity. The main function of the bone marrow is to supply marrow. Aspiration of the marrow has been utilized from mature hematopoietic cells for circulating blood in a steady- cytologic assessment, with analysis directed toward state as well as to respond to increased physiologic or morphology and obtainment of differential cell count. pathologic demands. “Romanowsky Stain”is a general term which includes Recent advances in the treatment of hematologic several specific stains or stain combinations. Romanowsky malignancies have been paralleled by renewed interest on (Giemsa & Leishman) stains are routinely used in 2 the part of pathologists and hematologists in methods of hematology obtaining and preparing bone marrow for diagnostic studies. In H&E stained sections of bone marrow, the more mature Bone marrow aspirate is a cytologic preparation of bone stages of the erythroid and myeloid cells, adipocytes and marrow cells obtained by aspiration of marrow and a smear megakaryocytes can be identified. However lymphoid cells of the cells. Bone marrow aspirate smears are primarily used as well as immature progenitor cells cannot be reliably 1 of 7 A Comparative Study Of Bone Marrow Aspiration Smear And Cell Block Using Romanwasky And Hematoxylin And Eosin Stain identified. The use of H&E and Romanowsky group of Other sites for Bone Marrow Aspiration4 are Posterior stains in the bone marrow analysis are very important and superior iliac spine, Posterior iliac crest, Anterior iliac crest, allow for studies of marrow’s overall cellularity, detection of Spinous process of lumber vertebra, Upper end of tibia. The focal lesions and extent of infiltration by various sternum remains the most commonly employed aspiration pathological entities. site. Bone marrow studies aid in the diagnosis, staging and Evaluation of Bone Marrow Aspiration Smears5: Detailed monitoring of several diseases Thus the present study was clinical history with clinical features like pallor, icterus, being conducted to compare efficacy & to explore further hepato or spleenomegaly and lymphadenopathy should be utility of Geimsa and Leishman stains of Romanowsky known. Peripheral blood films should be stained and group in bone marrow aspiration smears. evaluated. Stain 2 bone marrow smears with Leishman and Giemsa stain. Examined under scanner and low power, to OBJECTIVES asses cellularity, megakaryocyte, metastatic carcinoma cells The present study was undertaken with the objectives – which usually are present in the tail of the film. Select the area where cells are very well spread out. Bone marrow MATERIALS AND METHODS smears are ready for microscopic examination. The present study was done in Hamidia Hospital, Bhopal. Department of pathology from May 2008 to Sep. 2009. A CELL BLOCKS PREPARATION consecutive marrow samples from 83 patients attending The clot section is prepared from the blood left over after hematology OPD or ward of Hamidia Hospital, Bhopal. In aspirate smears have been made. The blood containing the present study included 83 cases in which complete admixed marrow particles is transferred to a container with peripheral smears, bone marrow and cell blocks was 1ml of 10% formal saline and 2ml methanol for fixation. available. The selection of cases was based on the clinical Future processing is carried out as for routine examination and peripheral blood smear of patients in which histopathological staining. a hematological disorder was suspected. The slides were viewed and reported. The reporting was Inclusion criteria were unexplained anemia, splenomegaly, done in conventional way as well as a scoring system created hepatomegaly and lymphadenopathy, Pancytopenia, indigenously leucopenia & leucocytosis, unexplained thrombocytopenia and thrombocytosis, Leukaemia, Myeloproliferative Figure 1 disorders, Lymphoproiferative disorders, Plasma cell dyscrasias. Exclusion criteria were the cases which were bleeding severely, like haemophilic patient and patient having platelet The maximum score for a single case, taking into account all count less than 10,000. the four parameters was 12. Thus the maximum possible History and clinical examination was done as per proforma. score in the study was calculated by multiplying the number Investigations like Hemoglobin, Total WBC count, Platelet of cases by 12 for each of 2 stains. A “quality index” was count, Reticulocyte count, Peripheral blood smear obtained by finding out the ratio of actual score obtained to examination, Blood glucose, Blood urea, Serum creatinine, the maximum score possible. AST, ALT, Bone marrow aspiration were performed Quality index = Actual score obtained / Maximum score Bone Marrow Aspiration was done through safe and possible. preferred sites. The preffered site for young and old patients nd rd The quality index for each of two stains was compared. As is Sternum at the level of the 2 or 3 interspace, just to the smears of all the cases were stained with Leishman stain one side of the midline, with the patient in a supine position. 3 and Geimsa stain, Quality Index of all slides were calculated Tibia for use is children younger that 1 year old the flat and compared. triangular area at the proximal end of the media surface of the tibia, just below the tibial tubercle was chosen. 2 of 7 A Comparative Study Of Bone Marrow Aspiration Smear And Cell Block Using Romanwasky And Hematoxylin And Eosin Stain OBSERVATIONS Figure 5 The present study was conducted for comparison between Table No.4: Result of scoring and Quality Index of Leishman and Giemsa stains 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- Figure 2 Table No.1: Various Hematological disorders Figure 6 Figure 3 Table No.5: Comparison of Quality Index in Leishman & Table No.2: Distribution of Heamatological Disorders Giemsa stained slides according to age Figure 4 Table No.3: Distribution of Hematological Disorders according to sex Figure 7 Table No.6: Various components seen in H & E stain blocks and Romanwsky stained bone marrow aspirate smears. 3 of 7 A Comparative Study Of Bone Marrow Aspiration Smear And Cell Block Using Romanwasky And Hematoxylin And Eosin Stain RESULTS Figure 9 By Romanowsky group of stains and Hematoxylin & Eosin Photo – 2: Giemsa stain (X400) Smudgy nuclear details 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
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