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Single-color Multitarget Flow Cytometry Using Monoclonal Antibodies Labeled with Different Intensities of the Same Fluorochrome
Annals of Laboratory Medicine ; : 171-176, 2012.
Artigo em Inglês | WPRIM | ID: wpr-80828
ABSTRACT

BACKGROUND:

We developed a single-color multitarget flow cytometry (SM-FC) assay, a single-tube assay with graded mean fluorescence intensities (MFIs). We evaluated the repeatability of SM-FC, and its correlation with multicolor flow cytometry (MFC), to assess its application as a routine FC assay.

METHODS:

We selected CD19, CD3, CD4, and CD8 as antigen targets to analyze a lymphocyte subset. MFIs were graded by adjusting monoclonal antibody (mAb) volumes to detect several cell populations. Dimly labeled mAb was prepared by decreasing mAb volume and the optimum diluted volume was determined by serial dilution. SM-FC repeatability was analyzed 10 times in 2 normal controls. The correlation between SM-FC and MFC was evaluated in 20 normal and 23 patient samples.

RESULTS:

CV values (0.8-5.0% and 1.3-4.1% in samples 1 and 2, respectively) acquired by SM-FC with CD3-fluorescein alpha-isothyocyanate (FITC)dim+CD4-FITCbright and with CD19-FITCdim+CD3-FITCbright showed good repeatability, comparable to that acquired by MFC (1.6-3.7% and 1.0-4.8% in samples 1 and 2, respectively). Excellent correlation was observed between the 2 methods in the 20 normal samples (B cells, T cells, non-Thelper cells, and Thelper cells; r2=0.87, 0.97, 0.97, and 0.98, respectively; P or =0.98, 0.99, 0.99, and 0.99, respectively; P<0.05).

CONCLUSIONS:

The multicolor, single-tube SM-FC technique is a potential alternative tool for identifying a lymphocyte subset.
Assuntos

Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Antígenos CD4 / Subpopulações de Linfócitos B / Subpopulações de Linfócitos T / Fluoresceína-5-Isotiocianato / Antígenos CD8 / Complexo CD3 / Cor / Antígenos CD19 / Citometria de Fluxo / Anticorpos Monoclonais Limite: Humanos Idioma: Inglês Revista: Annals of Laboratory Medicine Ano de publicação: 2012 Tipo de documento: Artigo

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Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Antígenos CD4 / Subpopulações de Linfócitos B / Subpopulações de Linfócitos T / Fluoresceína-5-Isotiocianato / Antígenos CD8 / Complexo CD3 / Cor / Antígenos CD19 / Citometria de Fluxo / Anticorpos Monoclonais Limite: Humanos Idioma: Inglês Revista: Annals of Laboratory Medicine Ano de publicação: 2012 Tipo de documento: Artigo