EurekaMag.com logo
+ Site Statistics
References:
53,869,633
Abstracts:
29,686,251
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on LinkedInFollow on LinkedIn

+ Translate

A novel cytolysis assay using fluorescent labeling and quantitative fluorescent scanning technology



A novel cytolysis assay using fluorescent labeling and quantitative fluorescent scanning technology



Journal of Immunological Methods 226(1-2): 29-41, June 24



A novel cellular cytotoxicity assay using Calcein acetoxymethyl (Calcein-AM), a cytoplasmic fluorescent label, has been developed as an alternative to the standard 51Chromium (Cr)-release. Target cells were loaded with Calcein-AM and then co-incubated with effector cells. An additional reagent, FluoroQuench, is added to extinguish fluorescence of dying target cells and of the culture media. Assay plates are read on a quantitative fluorescent scanner for determination of viable target cells. Percent lysis is calculated as one minus the percent viable cells as compared to fluorescent-labeled targets-only wells. The assay was tested to demonstrate the lytic activity of cytotoxic T lymphocyte (CTL) cultures, lymphokine-activated killer (LAK), and natural killer (NK) cell line effectors against peptide-pulsed and melanoma targets. In addition to the acquisition of results comparable to the 51Cr-release assay, the Calcein assay reliably measures cell-mediated cytotoxicity with little variance among replicates. The fluorescent assay represents a simple and useful alternative to the use of radioactive materials and adds the additional benefit of digital images and analysis.

(PDF emailed within 0-6 h: $19.90)

Accession: 010081503

Download citation: RISBibTeXText

PMID: 10410969

DOI: 10.1016/s0022-1759(99)00039-3



Related references

Scanning fluorescent microscopy is an alternative for quantitative fluorescent cell analysis. Cytometry. Part A 60(1): 53-62, 2004

Proprio spinal connections in cat studied by means of anterograde fluorescent labeling of nerve fibers combined with retrograde fluorescent labeling of moto neurons. Neuroscience Letters (SUPPL 14): S381, 1983

Rapid fluorescent labeling of tissue for microdissection using fluorescent specific binding agents. Official Gazette of the United States Patent & Trademark Office Patents 1286(2), Sep 14, 2004

Characterizaton of cells using non-fluorescent antibody labeling followed by fluorescent in situ hybridization. In Vitro Cellular & Developmental Biology Animal 35(3 PART 2): 28 A, March, 1999

A green-emitting fluorescent protein from Galaxeidae coral and its monomeric version for use in fluorescent labeling. Journal of Biological Chemistry 278(36): 34167-34171, 2003

A new assay for proteases using fluorescent labeling of proteins. Analytical Biochemistry 121(2): 290-294, 1982

Events in concanavalin a induced cytolysis of masto cytoma cells as seen by scanning electron microscopy and fluorescent microscopy. Israel Journal of Medical Sciences 13(3): 341-342, 1977

Probing functional interfaces of rod PDE gamma-subunit using scanning fluorescent labeling. Cell Biochemistry and Biophysics 28(2-3): 115-133, 1998

Site-specific incorporation of fluorescent probes into protein: hexahistidine-tag-mediated fluorescent labeling with (Ni(2+):nitrilotriacetic Acid (n)-fluorochrome conjugates. Journal of the American Chemical Society 123(48): 12123-5, 2001

Preparation and characterization of Fe3O4/CdTe magnetic/fluorescent nanocomposites and their applications in immuno-labeling and fluorescent imaging of cancer cells. Langmuir 26(2): 1278-1284, 2010

In Vivo Fluorescent Labeling of Tumor Cells with the HaloTagĀ® Technology. Current Chemical Genomics 6: 48-54, 2012

Fluorescent labeling of hormone receptors in viable cells preparation and properties of highly fluorescent derivatives of epidermal growth factor and insulin. Proceedings of the National Academy of Sciences of the United States of America 75(5): 2135-2139, 1978

Fluorescent nanoparticles as labels for immunometric assay of C-reactive protein using two-photon excitation assay technology. Analytical Biochemistry 328(2): 210-218, 2004

Arylacetamide-derived fluorescent probes: synthesis, biological evaluation, and direct fluorescent labeling of kappa opioid receptors in mouse microglial cells. Journal of Medicinal Chemistry 39(8): 1729-1735, 1996

Golgi like labeling of neurons with fluorescent dextrans visualized with confocal scanning laser microscopy. Society for Neuroscience Abstracts 17(1-2): 372, 1991