CORE
🇺🇦
make metadata, not war
Services
Services overview
Explore all CORE services
Access to raw data
API
Dataset
FastSync
Content discovery
Recommender
Discovery
OAI identifiers
OAI Resolver
Managing content
Dashboard
Bespoke contracts
Consultancy services
Support us
Support us
Membership
Sponsorship
Community governance
Advisory Board
Board of supporters
Research network
About
About us
Our mission
Team
Blog
FAQs
Contact us
Quantification of cells with specific phenotypes II: Determination of CD4 expression level on reconstituted lyophilized human PBMC labelled with anti‐CD4 FITC antibody
Authors
A.
A. K.
+45 more
Abbasi F.
B.
Braun
D.
D.
Dinesh K. Dikshit
Engel
G.
Gaigalas
J.
J.
J.
John
Kammel
Kim
Kuhne
L.
L.
L.
Lekishvili
M
M.
M.
M. and Neukammer
M..
M.O.
Marc-Olivier Baradez
Marshall
Marti
Ost
R.
R.
Revel
Roemer
S.
Sassi M
Schneider
Stebbings
Sutherland
T.
V.
Vonsky
Wang
Wang
Whitby
Publication date
1 January 2015
Publisher
'Wiley'
Doi
Cite
Abstract
This report focuses on the characterization of CD4 expression level in terms of equivalent number of reference fluorophores (ERF). Twelve different flow cytometer platforms across sixteen laboratories were utilized in this study. As a first step the participants were asked to calibrate the fluorescein isothiocyanate (FITC) channel of each flow cytometer using commercially available calibration standard consisting of five populations of microspheres. Each population had an assigned value of equivalent fluorescein fluorophores (EFF denotes a special case of the generic term ERF with FITC as the reference fluorophore). The EFF values were assigned at the National Institute of Standards and Technology (NIST). A surface-labelled lyophilized cell preparation was provided by the National Institute of Biological Standards and Control (NIBSC), using human peripheral blood mononuclear cells (PBMC) pre-labeled with a FITC conjugated anti-CD4 monoclonal antibody. Three PBMC sample vials, provided to each participant, were used for the CD4 expression analysis. The PBMC are purported to have a fixed number of surface CD4 receptors. On the basis of the microsphere calibration, the EFF value of the PBMC samples was measured to characterize the population average CD4 expression level of the PBMC preparations. Both the results of data analysis performed by each participant and the results of centralized analysis of all participants' raw data are reported. Centralized analysis gave a mean EFF value of 22,300 and an uncertainty of 750, corresponding to 3.3% (level of confidence 68%) of the mean EFF value. The next step will entail the measurement of the ERF values of the lyophilized PBMC stained with labels for other fluorescence channels. The ultimate goal is to show that lyophilized PBMC is a suitable biological reference cell material for multicolor flow cytometry and that it can be used to present multicolor flow cytometry measurements in terms of ABC (antibodies bound per cell) units. © 2015 International Society for Advancement of Cytometr
Similar works
Full text
Available Versions
Archivio istituzionale della ricerca - INRIM
See this paper in CORE
Go to the repository landing page
Download from data provider
oai:iris.inrim.it:11696/30250
Last time updated on 12/11/2016