✦ LIBER ✦

Simultaneous measurement of five-cell surface antigens by five-colour immunofluorescence

✍ Scribed by Andrew J. Beavis; Kenneth J. Pennline

Publisher: John Wiley and Sons
Year: 1994
Tongue: English
Weight: 682 KB
Volume: 15
Category: Article
ISSN: 0196-4763
DOI: 10.1002/cyto.990150413

No coin nor oath required. For personal study only.

✦ Synopsis

Multicolour immunofluorescence and flow cytometry were used for simultaneous measurement of five-cell surface antigens on murine spleen cells. We have been able to quantitate T-cells, T-cell subsets, B cells, and expression of the activation marker I-Ad from a single sample using four directly conjugated monoclonal antibodies LYT2-APC, L3T4-PE, B220-RED613, I-Ad-FITC and one indirect step THY 1.2-biotin/streptavidin-Cascade Blue. Three excitation wavelengths were used (488 nm, 647 nm, and U.V. 351-364 nm) for fluorescence measurements. The combination of fluorochromes used provided good resolution such that all five fluorescence signals were spectrally resolved. The percentage of cells positive for expression of a specific cell surface marker were almost identical for singlecolour samples and the five-colour analysis, differing by only 0.3-1.5 percentage points.

📜 SIMILAR VOLUMES

Enhanced immunofluorescence measurement

Enhanced immunofluorescence measurement resolution of surface antigens on highly autofluorescent, glutaraldehyde-fixed cells analyzed by phase-sensitive flow cytometry

✍ J.A. Steinkamp; N.M. Lehnert; J.F. Keij; B.E. Lehnert 📂 Article 📅 1999 🏛 John Wiley and Sons 🌐 English ⚖ 277 KB 👁 2 views

## Background: The primary source of interference in immunofluorescence measurements by flow cytometry is background autofluorescence. Methods: Using human lung fibroblasts (HLFs) as an autofluorescent cell model, unfixed HLFs and HLFs fixed in methanol, ethanol, formaldehyde, paraformaldehyde and

Simultaneous measurement of DNA content

Simultaneous measurement of DNA content and cell-surface immunofluorescence of human bone marrow cells using a single laser flow cytometer

✍ Paul P. T. Brons; Arie H. M. Pennings; Clemens Haanen; Hans M. C. Wessels; Jan B 📂 Article 📅 1990 🏛 John Wiley and Sons 🌐 English ⚖ 816 KB

## Abstract This paper describes the measurement of S phase DNA content in human bone marrow subpopulations using a single laser method for bivariate analysis of DNA content and cell‐surface immunofluorescence (s‐IF). Low density (<1.077 g/ml) bone marrow cells were labeled with a panel of unconjug

Detection and Measurement of a Single Bl

Detection and Measurement of a Single Blood Cell Surface Antigen by Thermal Lens Microscopy

✍ Hiroko Kimura; Fumiko Nagao; Asako Kitamura; Kazuya Sekiguchi; Takehiko Kitamori 📂 Article 📅 2000 🏛 Elsevier Science 🌐 English ⚖ 123 KB

A highly sensitive method for detection of antigens on the surface of a single blood cell using thermal lens microscopy is described. Colloidal gold, coated with antibody, was used to stain membrane antigens of leukocytes. Human leukocyte antigens on the lymphocytes and mononuclear leukocytes were o

Circulating CD4+ T lymphocytes with intr

Circulating CD4+ T lymphocytes with intracellular but no surface CD3 antigen in five of seven patients consecutively diagnosed with angioimmunoblastic T-cell lymphoma

✍ Stefan Serke; Antje van Lessen; Michael Hummel; Agnes Szczepek; Dieter Huhn; Har 📂 Article 📅 2000 🏛 John Wiley and Sons 🌐 English ⚖ 224 KB 👁 2 views

Optimization of a flow cytometric method

Optimization of a flow cytometric method for the simultaneous measurement of cell surface antigen, DNA content, and in vitro BrdUrd incorporation into normal and malignant hematopoietic cells

✍ Mette Holm; Mette Thomsen; Morten Høyer; Peter Hokland 📂 Article 📅 1998 🏛 John Wiley and Sons 🌐 English ⚖ 137 KB 👁 2 views

We have designed an assay for the simultaneous measurement of cell surface phenotype, S-phase fraction, and DNA content by single laser instrumentation for the purpose of determining the labeling index (LI), duration of S-phase (Ts), and the potential doubling time (Tpot) of leukocyte subpopulations