PMID: 8592577Jan 1, 1995Paper

Evaluation of different fixation-permeabilization methods for simultaneous detection of surface, cytoplasmic markers and DNA analysis by flow cytometry in some human hematopoietic cell lines

Neoplasma
M GlasováO Babusíková

Abstract

Different fixation/permeabilization methods were investigated for their convenience for simultaneous detection of membrane marker/DNA staining or cytoplasmic marker/DNA by flow cytometry. Nine different methods were employed. The expression of membrane marker CD20 and cytoplasmic marker CD22 on BJAB and DAUDI cells and cytoplasmic CD3 on MOLT4 cells were measured. Optimal methods were those that combine paraformaldehyde and saponin (for membrane CD20/DNA staining and cytoplasmic CD3/DNA staining) or buffered formaldehyde-acetone (for membrane CD20/DNA staining and for cytoplasmic CD22/DNA staining). Special interest was focused on proliferation marker CD71 and nuclear antigen Ki67. We investigated CD71 in MOLT4 cells and Ki67 in MOLT4, DAUDI and BJAB cells. Both of these markers are closely associated with proliferation rate. The optimal method for detection of Ki67/DNA staining combines paraformaldehyde with Tween 20.

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.