The role of flow cytometric immunophenotyping in improving the diagnostic accuracy in referred fine‐needle aspiration specimens

Abstract

Flow cytometric (FCM) immunophenotyping has an important role in the diagnostic work up of fine-needle aspiration (FNA) specimens obtained from lymphoid lesions. The objective of the present study was to evaluate the feasibility of this method with respect to referred FNA specimens. One hundred and two FNA specimens referred to our laboratory for FCM analysis during the last 3 years were studied. Specimens were sent, accompanied by cytological smears, from 11 distant hospitals by ordinary mail. The evaluation of potential B-cell monoclonality, the main diagnostic issue to be resolved using FCM, was possible in 86 of these 102 cases. The remaining 16 samples could not be analyzed or adequately interpreted because of sparse or necrotic material. A monoclonal B-cell population was found in 17/86 satisfactory cases, of which 16 were histologically confirmed. Eight cases contained cells positive for the epithelial marker Ber-EP4 and were diagnosed accordingly as carcinomas. FCM analysis of specimens obtained with a clinical question of Hodgkin lymphoma or T-cell lymphomas did not yield definitive data. The time lapse between sampling and analysis (12-84 hr) did not affect the results. This probably was due to the fact that all aspirates were taken in Roswell Park Memorial Institute (RPMI) cell medium, supplemented with 50% fetal calf serum. In conclusion, this retrospective study establishes that it is possible, in the majority of cases, to refer FNA material for FCM immunophenotyping by mail, and that results regarding B-cell clonality in the case of small-cell lymphomas are reliable also after a transportation period of 3-4 days. Carcinoma may be similarly diagnosed and a diagnosis of lymphoma may be excluded in reactive proliferations. Cases with only a few atypical cells or specimens from patients suspected of having Hodgkin lymphoma or T-cell lymphomas are not suitable for analysis by FCM.

Fine needle aspiration of abdominal organs: a review of current recommendations for achieving a diagnostic sample

“…A major advantage of the non-aspiration technique is reduced blood contamination when sampling highly vascular tissues, such as the spleen (Leblanc et al 2009 ), kidney and liver (Savage et al 1995, Raskin & Meyer 2015. Non-aspiration has been found to have similar or greater sensitivity to that of aspiration for a variety of tissues, including the spleen and abdominal lymph nodes (Menard & Papageorges 1995, Savage et al 1995, Wallace et al 2001, Sigstad et al 2004, Leblanc et al 2009, Sajeev & Siddaraju 2009, Jahromi et al 2015. In contrast, other studies have shown an increased diagnostic yield in the liver (Haseler et al 2011 ) and pancreas (Kinney et al 1993 ) when using aspiration as opposed to nonaspiration techniques.…”

“…Flow cytometry can be performed on FNA specimens and has been shown to be an accurate method for classification of cell lineage for abdominal organ samples in lymphoma patients (Lastra et al 2015, Guzera et al 2016. However, this method requires at least 6×10 6 cells to be mixed with serum or cell preservative and placed in a plain serum tube, and is ideally analysed within 24 hours of collection, although satisfactory results have been achieved in human samples that were several days old (Sigstad et al 2004, Colorado State University 2016a, b , Guzera et al 2016.…”

Basics of cytology

“…This means that after the initial evaluation of the smears the pathologist/cytopathologist will decide if additional material is needed to do ancillary studies such as cultures, molecular pathology studies, cytogenetic analysis, and immunophenotypic analysis by flow cytometry. [87–89]…”

Importance of Flow Cytometry in the Cytopathologic Evaluation of Lymphoid Lesions Involving the Kidney

“…Newer techniques such as immunophenotyping, cytogenetics and molecular studies, however, have opened promising avenues for the initial diagnosis of NHL on small samples [7,8,9,10,11], and numerous studies performed over the past 20 years have shown FNA to have both a high sensitivity and specificity in the diagnosis of NHL when used with ancillary techniques [12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29]. When FNA cytomorphology is combined with FC on a sample from any organ and the findings are compared with follow-up surgical specimens, diagnostic sensitivity and specificity have been reported as high as 97 and 94%, respectively [6].…”