Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Egger, G; Burda, A; Glasner, A.
A simple method for measuring the F-actin content of human polymorphonuclear leukocytes in whole blood.
Virchows Arch. 2001; 438(4):394-397
Doi: 10.1007%2Fs004280000321
Web of Science
PubMed
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- Führende Autor*innen der Med Uni Graz
- Egger Gerd
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- Abstract:
- We have developed an improved method for measuring the filamentous (F) actin content of human blood polymorphonuclear leukocytes (PMNs). The essential feature of the method is the immediate fixation of the F-actin cytoskeleton. Fresh whole blood (100 microliters) is shock-cooled by the addition of 1.0 ml of a mixture of 18.75% glycerol and 5% formaldehyde in phosphate buffer pre-cooled to -8 degrees C and subsequently fixed at 4 degrees C for 15 min. After lysis in distilled water and removal of the red blood cells by centrifugation, the F-actin cytoskeleton of the PMNs is stained with fluorescein isothiocyanate (FITC)-phalloidin and quantified by means of flow cytometry. In healthy test subjects, PMN stimulation by the chemotactic peptide formyl-methionyl-leucyl-phenylalanine (FMLP) for 20 s resulted in a significantly increased F-actin assembly, while in patients with multiple organ failure, two subpopulations arose: one with greater F-actin content and a second with lower F-actin content in comparison with the unstimulated blood sample. This simple and fast method may be a useful tool in basic and clinical research.
- Find related publications in this database (using NLM MeSH Indexing)
- Actins - blood
- Adult - blood
- Cytoskeleton - chemistry
- Female - chemistry
- Flow Cytometry - methods
- Fluorescein-5-isothiocyanate - chemistry
- Humans - chemistry
- Male - chemistry
- Multiple Organ Failure - blood
- N-Formylmethionine Leucyl-Phenylalanine - pharmacology
- Neutrophils - chemistry
- Phalloidine - chemistry
- Reference Values - chemistry
- Staining and Labeling - chemistry
- Find related publications in this database (Keywords)
- polymorphonuclear leukocytes
- PMN
- cytoskeleton
- actin
- FMLP