Preparation of bronchoalveolar lavage fluid with microscope slide smears

Abstract

The method of preparation of bronchoalveolar lavage fluid (BALF) for cytological examination can significantly affect the results of cellular quantitation. Investigations have shown that cytocentrifugation leads to an underestimation of the number of lymphocytes and membrane filter preparation to an underestimation of the number of neutrophils. As a simple alternative to these two techniques, BALF cells could be prepared by the microscope slide smear technique, which is familiar as the means for preparing peripheral blood for differential counts.In order to compare cell differentials determined by microscope slide technique with differentials resulting from cytocentrifugation, cells were isolated from 35 BALF samples using standard methods, and counted using a haematocytometer. Forty thousand cells in 200 µL were prepared by cytocentrifugation (3 min, 57×g; Cytospin 2) and 5×10 5 cells in 5 µL by microscope slide smear. Both samples were air-dried, stained using May-Grünwald Giemsa stain, and 600 cells were counted to obtain differentials. To test the adequacy of sampling by the microscope slide smear technique, known quantities of lymphocytes or neutrophils were added to fixed numbers of BALF cells, microscope slide smears prepared, and differentials determined on 600 cells. The resulting differentials were compared to the calculated differentials.Preparation Thus, microscope slide smear preparation is a simple and accurate method for the quantitation of bronchoalveolar lavage fluid cytology.

Pediatric Bronchoalveolar Lavage: Practical Considerations and Future Prospects

“…Once the slide has been made, a WBC differential count is performed, and the number of cells counted recommended to achieve this is also variable (300-600) [19,23]. If cytocentrifugation is used to prepare slides for differential evaluation, a minimum of 300 cells is recommended to reliably enumerate the cellularity of the sample [24].…”

Comparação entre as técnicas de esfregaço linear e citocentrifugação para avaliação citológica do lavado broncoalveolar equino

“…Não foram encontradas alterações morfológicas significativas e nem diferenças estatísticas entre nenhum dos tipos celulares processados pelos dois métodos, o que permite afirmar que o método de esfregaço linear é uma alternativa segura para avaliação morfológica celular do LBA de equinos, podendo ser utilizado no lugar da citocentrifugação quando esta não estiver disponível. Cytological characteristics observed in the linear smear were less preserved than in the cytocentrifugation method, as previously observed 7,8 , but BAL cells prepared by the linear smear technique yielded well preserved cell morphology, as observed by Thomp-son et al 6 . Though the cytocentrifugation technique is the method of choice 6,7,8 , the linear smear is a reliable alternative for cytological analyses of equine BALs and can be recommended as a substitution to cytocentrifugation when the latter cannot be applied routinely.…”

Overexpression of Bcl-2 Protein in Bronchoalveolar Lavage Lymphocytes and Macrophages in Sarcoidosis

“…Fiberoptic bronchoscopy with BAL was performed with the informed consent of the patient as part of routine clinical management, according to recommended guidelines and previous reports [13, 14]. Patients with a forced expiratory volume in 1 s (FEV 1 ) <1 liter and PaO 2 <60 mm Hg were excluded from BAL.…”

“…BALF cell counts were completed as previously reported [13, 14]. Total cell counts were determined using an improved Neubauer counting chamber and expressed as the total number of cells per milliliter of aspirated fluid.…”