Single-Tube Flow Cytometry Assay for the Detection of Mature Lymphoid Neoplasms in Paucicellular Samples

Abstract

Objectives: Flow cytometry (FC) has become a useful support for cytomorphologic evaluation (CM) of fine-needle aspirates (FNA) and serous cavity effusions (SCE) in cases of suspected non-Hodgkin lymphoma (NHL). FC results may be hampered by the scarce viability and low cellularity of the specimens. Study Design: We developed a single-tube FC assay (STA) that included 10 antibodies cocktailed in 8-color labeling, a cell viability dye, and a logical gating strategy to detect NHL in hypocellular samples. The results were correlated with CM and confirmed by histologic or molecular data when available. Results: UsingtheSTA, we detected B-type NHL in 31 out of 103 hypocellular samples (81 FNA and 22 SCE). Of these, 8 were not confirmed by CM and 2 were considered to be only suspicious. The FC-negative samples had a final diagnosis of benign/reactive process (42/72), carcinoma (27/72), or Hodgkin lymphoma (3/72). Conclusions: The STA approach allowed obtainment of maximum immunophenotyping data in specimens containing a low number of cells and a large amount of debris. The information obtained by STA can help cytomorphologists not only to recognize but also to exclude malignant lymphomas.

Flow cytometry in reactive lymph node aspirates: Evaluation of distribution of populations with immunophenotypic outliers

“…Stacchini et al 7 observed CD4:CD8 ratios with ranges of 0.1‐1.7 and 0.8‐4.4 in cases of reactive hyperplasia and metastasis, respectively. Three cases of Hodgkin lymphoma exhibited a CD4:CD8 ratio ranging from 3.9‐7.3 in their study.…”

‘Six Memos' for Lymph Node Fine-Needle Cytology and Flow Cytometry

“…Numerous studies showed that FNC is suitable for different molecular procedures and LN FNC-FC plays a key role in guaranteeing the best management of the available material. The article by Peluso et al [19] describes the most common molecular procedures used for NHL and their potential application on FNC material, and the article by Stacchini et al [20] focuses on small samples and specific technical solutions. In conclusion, the six features highlighted by Calvino identify qualities shared by FC, which can be fully exploited in the diagnosis of LN-FNC and represent the basis for the application of specific molecular procedures.…”