Volumetric capillary cytometry: a new method for absolute cell enumeration

Cytometry
L J DietzN L Sizto

Abstract

This article describes a novel cytometry system that is capable of imaging, identifying, and counting populations of fluorescently labeled cells within a precisely known volume of unlysed whole blood contained in a precision capillary. The whole blood is reacted with dye-labeled antibodies directed at cell surface antigens. A red excitation source (helium-neon laser) is used to minimize auto-fluorescence and interference by blood components. A two-color fluorescence detection system is described, using the dyes Cy5 and Cy5.5, allowing the simultaneous detection and quantitation of two different types of surface antigens. Image processing techniques are used to identify cellular events in the image and to quantitate the fluorescent intensities in both color planes. The utility of this technique is demonstrated by performing absolute CD3+/CD4+ lymphocyte counts in whole blood, with minimal sample preparation, and with the blood sample completely contained in a disposable cartridge. A correlation study comparing this technique to flow cytometry gives a correlation coefficient of 0.98. In addition, the ability of this system to monitor reaction kinetics is demonstrated with a measurement of the rate of association of the anti-CD4 a...Continue Reading

Citations

Feb 8, 2000·Analytical Biochemistry·L C Taylor, D R Walt
Sep 29, 1999·Proceedings of the National Academy of Sciences of the United States of America·P ZuckJ Inglese
Jan 12, 2010·Journal of Biomedical Optics·Homanaz GhafariQuentin S Hanley
Jul 11, 2014·ACS Chemical Biology·Francisco CiruelaVíctor Fernández-Dueñas
Aug 4, 2004·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Howard M Shapiro
May 9, 2006·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Howard M Shapiro, Nancy G Perlmutter
Nov 1, 2006·Allergology International : Official Journal of the Japanese Society of Allergology·Eiji MatsukumaNaomi Kondo
Mar 14, 2002·Cytometry·Arjan G J TibbeLeon W M M Terstappen
Feb 9, 2002·Journal of Clinical Apheresis·N D Depcik-SmithM E Brecher
Dec 10, 1999·Cytometry·I L BarbosaA Carvalhais
Jun 29, 2011·Journal of Acquired Immune Deficiency Syndromes : JAIDS·Kasama SukapiromKovit Pattanapanyasat
Jan 9, 2009·Cytometry. Part B, Clinical Cytometry·Sook Young BaeChae Seung Lim
Jan 26, 2016·Cytometry. Part B, Clinical Cytometry·Morgane GossezGuillaume Monneret
Dec 8, 2006·Biomedical Microdevices·An-Shik Yang, Wen-Hsin Hsieh
Dec 10, 1999·Nature Biotechnology·A G TibbeL W Terstappen
Feb 9, 2007·Clinical and Vaccine Immunology : CVI·Xiao LiJan Greve
Sep 5, 2008·Current Protocols in Cytometry·Howard M Shapiro
Feb 22, 2011·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Tycho M ScholtensLeon W M M Terstappen
Mar 25, 2004·BioTechniques·Aaron B KantorLouis J Dietz
Sep 23, 2000·Journal of Microbiological Methods·H M Shapiro

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.