Comparative analysis of three permeabilization methods for cytofluorometric evaluation of cytoplasmic myeloperoxidase.

S. P. Tay, S. K. Cheong, Noor Hamidah Hussin, O. Ainoon

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

A comparative study was conducted to evaluate three different permeabilization methods: FACS Permeabilizing Solution (FPerm), CytoFix/CytoPerm Kit (CFP) and Paraformaldehyde-Tween 20 (PFT) reagents, in cytoplasmic labeling of myeloperoxidase (MPO). Peripheral blood cells from 23 healthy subjects were fixed and permeabilized according to the proposed procedures, prior to direct immunofluorescence staining with CD14, CD45, IgG1, IgG2 and MPO monoclonal antibodies (McAb). Subsequent flow cytometric analysis was performed on FACSCalibur flow cytometer (Becton Dickinson, BD). As far as the antigenic expression of MPO in normal samples is concerned, FPerm and CFP demonstrated better cytoplasmic staining by inducing minor effects on light-scattering properties of the cell populations, whereas PFT-treated samples showed a diminished ability to distinguish the cell types. However, the simple and rapid FPerm method required an earlier processing of samples since the stored whole blood samples (for more than 8 hours) tended to show a significant decrease of fluorescence intensity. We also have demonstrated that P/N ratio possesses added value in evaluation of cell reactivity in immunophenotyping, based upon the apparent nonspecific cytoplasmic staining of MPO in the lymphocyte population.

Original languageEnglish
Pages (from-to)37-43
Number of pages7
JournalThe Malaysian journal of pathology
Volume21
Issue number1
Publication statusPublished - 1999

Fingerprint

Peroxidase
Polysorbates
Staining and Labeling
Immunoglobulin G
Immunophenotyping
Direct Fluorescent Antibody Technique
Population
Blood Cells
Healthy Volunteers
Fluorescence
Monoclonal Antibodies
Lymphocytes
Light
paraform

Cite this

Comparative analysis of three permeabilization methods for cytofluorometric evaluation of cytoplasmic myeloperoxidase. / Tay, S. P.; Cheong, S. K.; Hussin, Noor Hamidah; Ainoon, O.

In: The Malaysian journal of pathology, Vol. 21, No. 1, 1999, p. 37-43.

Research output: Contribution to journalArticle

@article{2dde9606f83e4add953b845dabe91c8f,
title = "Comparative analysis of three permeabilization methods for cytofluorometric evaluation of cytoplasmic myeloperoxidase.",
abstract = "A comparative study was conducted to evaluate three different permeabilization methods: FACS Permeabilizing Solution (FPerm), CytoFix/CytoPerm Kit (CFP) and Paraformaldehyde-Tween 20 (PFT) reagents, in cytoplasmic labeling of myeloperoxidase (MPO). Peripheral blood cells from 23 healthy subjects were fixed and permeabilized according to the proposed procedures, prior to direct immunofluorescence staining with CD14, CD45, IgG1, IgG2 and MPO monoclonal antibodies (McAb). Subsequent flow cytometric analysis was performed on FACSCalibur flow cytometer (Becton Dickinson, BD). As far as the antigenic expression of MPO in normal samples is concerned, FPerm and CFP demonstrated better cytoplasmic staining by inducing minor effects on light-scattering properties of the cell populations, whereas PFT-treated samples showed a diminished ability to distinguish the cell types. However, the simple and rapid FPerm method required an earlier processing of samples since the stored whole blood samples (for more than 8 hours) tended to show a significant decrease of fluorescence intensity. We also have demonstrated that P/N ratio possesses added value in evaluation of cell reactivity in immunophenotyping, based upon the apparent nonspecific cytoplasmic staining of MPO in the lymphocyte population.",
author = "Tay, {S. P.} and Cheong, {S. K.} and Hussin, {Noor Hamidah} and O. Ainoon",
year = "1999",
language = "English",
volume = "21",
pages = "37--43",
journal = "Malaysian Journal of Pathology",
issn = "0126-8635",
publisher = "Malaysian Society of Pathologists",
number = "1",

}

TY - JOUR

T1 - Comparative analysis of three permeabilization methods for cytofluorometric evaluation of cytoplasmic myeloperoxidase.

AU - Tay, S. P.

AU - Cheong, S. K.

AU - Hussin, Noor Hamidah

AU - Ainoon, O.

PY - 1999

Y1 - 1999

N2 - A comparative study was conducted to evaluate three different permeabilization methods: FACS Permeabilizing Solution (FPerm), CytoFix/CytoPerm Kit (CFP) and Paraformaldehyde-Tween 20 (PFT) reagents, in cytoplasmic labeling of myeloperoxidase (MPO). Peripheral blood cells from 23 healthy subjects were fixed and permeabilized according to the proposed procedures, prior to direct immunofluorescence staining with CD14, CD45, IgG1, IgG2 and MPO monoclonal antibodies (McAb). Subsequent flow cytometric analysis was performed on FACSCalibur flow cytometer (Becton Dickinson, BD). As far as the antigenic expression of MPO in normal samples is concerned, FPerm and CFP demonstrated better cytoplasmic staining by inducing minor effects on light-scattering properties of the cell populations, whereas PFT-treated samples showed a diminished ability to distinguish the cell types. However, the simple and rapid FPerm method required an earlier processing of samples since the stored whole blood samples (for more than 8 hours) tended to show a significant decrease of fluorescence intensity. We also have demonstrated that P/N ratio possesses added value in evaluation of cell reactivity in immunophenotyping, based upon the apparent nonspecific cytoplasmic staining of MPO in the lymphocyte population.

AB - A comparative study was conducted to evaluate three different permeabilization methods: FACS Permeabilizing Solution (FPerm), CytoFix/CytoPerm Kit (CFP) and Paraformaldehyde-Tween 20 (PFT) reagents, in cytoplasmic labeling of myeloperoxidase (MPO). Peripheral blood cells from 23 healthy subjects were fixed and permeabilized according to the proposed procedures, prior to direct immunofluorescence staining with CD14, CD45, IgG1, IgG2 and MPO monoclonal antibodies (McAb). Subsequent flow cytometric analysis was performed on FACSCalibur flow cytometer (Becton Dickinson, BD). As far as the antigenic expression of MPO in normal samples is concerned, FPerm and CFP demonstrated better cytoplasmic staining by inducing minor effects on light-scattering properties of the cell populations, whereas PFT-treated samples showed a diminished ability to distinguish the cell types. However, the simple and rapid FPerm method required an earlier processing of samples since the stored whole blood samples (for more than 8 hours) tended to show a significant decrease of fluorescence intensity. We also have demonstrated that P/N ratio possesses added value in evaluation of cell reactivity in immunophenotyping, based upon the apparent nonspecific cytoplasmic staining of MPO in the lymphocyte population.

UR - http://www.scopus.com/inward/record.url?scp=0033139783&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033139783&partnerID=8YFLogxK

M3 - Article

VL - 21

SP - 37

EP - 43

JO - Malaysian Journal of Pathology

JF - Malaysian Journal of Pathology

SN - 0126-8635

IS - 1

ER -