Immunophenotyping of Paucicellular Samples

Stacchini, Alessandra; Demurtas, Anna; Aliberti, Sabrina

doi:10.1002/0471142956.cy0946s68

Cited by 10 publications

(15 citation statements)

References 24 publications

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“…The overall rationale of the analysis and of the gating strategy applied has been previously published [11]. Examples of the STA protocol with the different cytometers utilized in this study are showed in figures 1 and 2.…”

Section: Resultsmentioning

confidence: 99%

“…Therefore, the set-up of a single tube, based on patient information and clinical suspicion, is frequent in FC laboratory practice. In this study, we report our experience with an FC assay that was developed in order to gather the greatest amount of information from hypocellular samples sent for lymphoma suspicion or monitoring [11]. The method used different MoAbs cocktailed in one tube to detect and identify lymphocytes, as well as a logical gating strategy to gate out dead cells and get rid of debris and clumped cells (doublets).…”

Section: Discussionmentioning

confidence: 99%

“…FNA were placed in Vacutainer lithium heparin tubes (BD Biosciences, San Jose, Calif., USA) containing RPMI 1640 with 10% fetal calf serum (holding medium); fluids were collected directly into Vacutainer lithium heparin tubes. All detailed procedures for FNA collection and cell processing have been previously published [4,11]. …”

Section: Methodsmentioning

confidence: 99%

“…FC investigations may, however, be hindered by the low cell number, limiting the number of tests that can be performed. Tailored panels and appropriate sample processing are mandatory in such cases [11]. This study reports our experience with paucicellular samples using an FC approach that employs a nuclear dye to recognize viable cells, antibodies cocktailed in 8-color labeling to detect lymphomas, and a Boolean gating strategy to analyze data.…”

Section: Introductionmentioning

confidence: 99%

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

Stacchini

Demurtas²,

Aliberti³

et al. 2016

Acta Cytologica

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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.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

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

Stacchini

Demurtas²,

Aliberti³

et al. 2016

Acta Cytologica

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“…The use of fluids and aspirates may entail a very limited number of cells and only a single tube to run FC investigations. In these cases, tailored antibody panels should be designed on the basis of clinical features, patient history and specimen source [13]. …”

Section: Fc Specimen Sourcementioning

confidence: 99%

Extranodal Lymphoproliferative Processes and Flow Cytometry

Stacchini¹,

Demurtas²,

Aliberti³

2016

Acta Cytologica

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Objective: Fine-needle aspiration (FNA) cytology is a safe and cost-effective technique for the diagnosis of lymphoproliferative processes, especially when correlated with clinical and imaging studies. However, cytology alone may be unable to detect a lymphoid neoplastic process, as architectural features are less obvious than in histologic preparations and, in certain cases, reactive processes may mimic lymphoma. Flow cytometry (FC) has been recognized as an important ancillary technique in the diagnosis of lymphoid neoplasms and it can be used in conjunction with FNA in the evaluation of lymphoproliferative processes. Study Design: We performed a review of the published literature concerning FC applied to the detection of salivary glands and thyroid lymphoproliferative processes, which are frequently related to autoimmune diseases and difficult to diagnose by cytomorphology alone. Results: FC is able to detect and subtype non-Hodgkin lymphomas and may contribute to the exclusion of a neoplastic process in cytologically unclear cases. Conclusions: FC can be successfully applied in the differential diagnosis of lymphoproliferative processes in the head and neck region. The FNA-FC combined approach can reduce time to therapy and may prevent unnecessary surgical biopsies.

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Method for DNA Ploidy Analysis Along with Immunophenotyping for Rare Populations in a Sample using FxCycle Violet

et al. 2017

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The clinical use of flow cytometric DNA ploidy assay has been extended towards stratifying the risk of diseases, such as monoclonal gammopathies or B cell acute lymphoblastic leukemia, and to detect circulating tumor cells, both of which require detection of minute cell populations. This unit describes a protocol for determining DNA ploidy in fixed samples with simultaneous surface immunophenotyping. It is an easy method for simultaneous 6- to 8-color immunophenotyping and DNA content analysis using FxCycle Violet (FCV; DAPI) dye. This protocol is a one-step modification of routine multicolor immunophenotyping that includes surface staining followed by fixation and then DNA staining with FCV. It utilizes mature lymphocytes from the sample as an internal control for determination of DNA index. It is a sensitive method that allows DNA-ploidy determination and cell cycle analysis in a rare tumor population as low as 100 events, as well as DNA ploidy determination in various subsets of hematopoietic cells in the same sample based on their immunophenotype. © 2017 by John Wiley & Sons, Inc.

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Immunophenotyping of Paucicellular Samples

Cited by 10 publications

References 24 publications

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

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

Extranodal Lymphoproliferative Processes and Flow Cytometry

Method for DNA Ploidy Analysis Along with Immunophenotyping for Rare Populations in a Sample using FxCycle Violet

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