PubMed Health⌕ Search
Home/Archive/PubMed

PubMed · 3283509

The microscopic double immunofluorescence technique, a method for quantitative differentiation between extra- and intracellularly located bacteria in isolated polymorphonuclear granulocytes.

L Dziwisch·J Heesemann·B Kremer
View original source ↗Export metadata ↓

Abstract

A simple microscopic double immunofluorescence staining technique for the study of phagocytosis of living bacteria by human polymorphonuclear granulocytes (PMN) is presented. This technique enables the differentiation between cell-attached and ingested bacteria and allows the kinetics of phagocytosis to be monitored. The practicability and accuracy of this method is demonstrated by using two genetically defined Yersinia enterocolitica strains of different virulence, and isolated PMN resuspended in serum-free and serum-containing medium, respectively. The advantages of this method over other techniques are discussed.

Explore related subjects

Fluorescent Antibody TechniqueGranulocytesHumansIn Vitro TechniquesPhagocytosisVirulenceYersinia enterocoliticaJournal Article

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

L Dziwisch, J Heesemann, B Kremer. 1988. The microscopic double immunofluorescence technique, a method for quantitative differentiation between extra- and intracellularly located bacteria in isolated polymorphonuclear granulocytes.. https://doi.org/10.1007/bf00189531

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

PubMed2007

The use of siRNA to validate immunofluorescence studies.

Cellular immunofluorescence studies can be validated by using either specific small interfering RNA (siRNA) duplexes or expression plasmids that induce the expression of specific siRNAs. The usage of either siRNA tool reduces the expression of the specific protein being studied, thus reducing substantially or abolishing the immunofluorescence detected when using a fluorescent antibody that recognizes the protein.

Fluorescent Antibody Technique↗
PubMed2006

Translational diffusion of fluorescent proteins by molecular fourier imaging correlation spectroscopy.

The ability to noninvasively observe translational diffusion of proteins and protein complexes is important to many biophysical problems. We report high signal/noise (>or=250) measurements of the translational diffusion in viscous solution of the fluorescent protein, DsRed. This is carried out using a new technique: molecular Fourier imaging correlation spectroscopy (M-FICS). M-FICS is an interferometric method that detects a collective Fourier component of the fluctuating density of a small population of fluorescent molecules, and provides information about the distribution of molecular diffusivities. A theoretical analysis is presented that expresses the detected signal fluctuations in terms of the relevant time-correlation functions for molecular translational diffusion. Furthermore, the role played by optical orientational degrees of freedom is established. We report Fickian self-diffusion of the DsRed tetramer at short timescales. The long-time deviation of our data from Fickian behavior is used to determine the variance of the distribution of the protein self-diffusion coefficient. We compare our results to the expected outcomes for 1), a bi-disperse distribution of protein species, and 2), dynamic disorder of the host solvent.

Fluorescent Antibody Technique↗