Micro-scale flow cytometry-based and biochemical analysis of lipid signaling in primary B cell subpopulations

Pierre Antony, Kristen Hoek, Bhaskarjyoti Sarmah, Wasif Khan

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

B cell subpopulations in the spleen have been extensively characterized phenotypically; however, biochemical properties of these cell populations following B cell antigen receptor engagement have not been fully determined due to technical difficulties and limiting cell numbers. We therefore employed mini-scale protocols to assess lipid signaling, particularly that of diacylglycerol and inositol trisphosphate, with as few as 0.5×106 purified early (T1) and late (T2) transitional B cells. Additionally, utilizing flow cytometric techniques, we determined levels of phosphatidylinositol bisphosphate and calcium mobilization in T1 and T2 cells, as well as mature follicular and marginal zone B cells using less than 1×106 primary B cells. Thus, these biochemical and flow cytometric methodologies can be used to analyse signal-induced changes in phosphatidylinositol bisphosphate levels, diacylglycerol and inositol triphosphate production and calcium in each B cell population.

Original languageEnglish
Pages (from-to)73-83
Number of pages11
JournalBiological Procedures Online
Volume9
Issue number1
DOIs
StatePublished - Dec 1 2007
Externally publishedYes

Fingerprint

Flow cytometry
Flow Cytometry
B-Lymphocytes
Cells
Lipids
Diglycerides
Inositol
Phosphatidylinositols
B-Cell Antigen Receptors
B-Lymphoid Precursor Cells
Population
Spleen
Cell Count
Calcium

Keywords

  • Antigen
  • B-cell
  • Immunomagnetic separation
  • Receptors

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Micro-scale flow cytometry-based and biochemical analysis of lipid signaling in primary B cell subpopulations. / Antony, Pierre; Hoek, Kristen; Sarmah, Bhaskarjyoti; Khan, Wasif.

In: Biological Procedures Online, Vol. 9, No. 1, 01.12.2007, p. 73-83.

Research output: Contribution to journalArticle

@article{981cfdaedd494bbfb3b00da045fb422d,
title = "Micro-scale flow cytometry-based and biochemical analysis of lipid signaling in primary B cell subpopulations",
abstract = "B cell subpopulations in the spleen have been extensively characterized phenotypically; however, biochemical properties of these cell populations following B cell antigen receptor engagement have not been fully determined due to technical difficulties and limiting cell numbers. We therefore employed mini-scale protocols to assess lipid signaling, particularly that of diacylglycerol and inositol trisphosphate, with as few as 0.5×106 purified early (T1) and late (T2) transitional B cells. Additionally, utilizing flow cytometric techniques, we determined levels of phosphatidylinositol bisphosphate and calcium mobilization in T1 and T2 cells, as well as mature follicular and marginal zone B cells using less than 1×106 primary B cells. Thus, these biochemical and flow cytometric methodologies can be used to analyse signal-induced changes in phosphatidylinositol bisphosphate levels, diacylglycerol and inositol triphosphate production and calcium in each B cell population.",
keywords = "Antigen, B-cell, Immunomagnetic separation, Receptors",
author = "Pierre Antony and Kristen Hoek and Bhaskarjyoti Sarmah and Wasif Khan",
year = "2007",
month = "12",
day = "1",
doi = "10.1251/bpo135",
language = "English",
volume = "9",
pages = "73--83",
journal = "Biological Procedures Online",
issn = "1480-9222",
publisher = "BioMed Central",
number = "1",

}

TY - JOUR

T1 - Micro-scale flow cytometry-based and biochemical analysis of lipid signaling in primary B cell subpopulations

AU - Antony, Pierre

AU - Hoek, Kristen

AU - Sarmah, Bhaskarjyoti

AU - Khan, Wasif

PY - 2007/12/1

Y1 - 2007/12/1

N2 - B cell subpopulations in the spleen have been extensively characterized phenotypically; however, biochemical properties of these cell populations following B cell antigen receptor engagement have not been fully determined due to technical difficulties and limiting cell numbers. We therefore employed mini-scale protocols to assess lipid signaling, particularly that of diacylglycerol and inositol trisphosphate, with as few as 0.5×106 purified early (T1) and late (T2) transitional B cells. Additionally, utilizing flow cytometric techniques, we determined levels of phosphatidylinositol bisphosphate and calcium mobilization in T1 and T2 cells, as well as mature follicular and marginal zone B cells using less than 1×106 primary B cells. Thus, these biochemical and flow cytometric methodologies can be used to analyse signal-induced changes in phosphatidylinositol bisphosphate levels, diacylglycerol and inositol triphosphate production and calcium in each B cell population.

AB - B cell subpopulations in the spleen have been extensively characterized phenotypically; however, biochemical properties of these cell populations following B cell antigen receptor engagement have not been fully determined due to technical difficulties and limiting cell numbers. We therefore employed mini-scale protocols to assess lipid signaling, particularly that of diacylglycerol and inositol trisphosphate, with as few as 0.5×106 purified early (T1) and late (T2) transitional B cells. Additionally, utilizing flow cytometric techniques, we determined levels of phosphatidylinositol bisphosphate and calcium mobilization in T1 and T2 cells, as well as mature follicular and marginal zone B cells using less than 1×106 primary B cells. Thus, these biochemical and flow cytometric methodologies can be used to analyse signal-induced changes in phosphatidylinositol bisphosphate levels, diacylglycerol and inositol triphosphate production and calcium in each B cell population.

KW - Antigen

KW - B-cell

KW - Immunomagnetic separation

KW - Receptors

UR - http://www.scopus.com/inward/record.url?scp=37649014499&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=37649014499&partnerID=8YFLogxK

U2 - 10.1251/bpo135

DO - 10.1251/bpo135

M3 - Article

C2 - 18385809

AN - SCOPUS:37649014499

VL - 9

SP - 73

EP - 83

JO - Biological Procedures Online

JF - Biological Procedures Online

SN - 1480-9222

IS - 1

ER -