A flow-cytometric gram-staining technique for milk-associated bacteria.
AuthorsHolm C, Jespersen L
JournalAppl Environ Microbiol
PubMed ID12732558
'A Gram-staining technique combining staining with two fluorescent stains, Oregon Green-conjugated wheat germ agglutinin (WGA) and hexidium iodide (HI) followed by flow-cytometric detection is described. WGA stains gram-positive bacteria while HI binds to the DNA of all bacteria after permeabilization by EDTA and incubation at 50 degrees C for 15 ... More
A fluorescent Gram stain for flow cytometry and epifluorescence microscopy.
AuthorsMason DJ, Shanmuganathan S, Mortimer FC, Gant VA
JournalAppl Environ Microbiol
PubMed ID9647848
'The fluorescent nucleic acid binding dyes hexidium iodide (HI) and SYTO 13 were used in combination as a Gram stain for unfixed organisms in suspension. HI penetrated gram-positive but not gram-negative organisms, whereas SYTO 13 penetrated both. When the dyes were used together, gram-negative organisms were rendered green fluorescent by ... More
Metabolic activity in filamentous fungi can be analysed by flow cytometry.
AuthorsBradner JR, Nevalainen KM
JournalJ Microbiol Methods
PubMed ID12782375
The use of flow cytometry in combination with fluorescent dyes as a technique to rapidly differentiate and enumerate bacterial and yeast cells is well established. We have shown that through the judicial choice of stains, the nondestructive screening and sorting of fungal material is possible. The early stages of growth, ... More
Increased sensitivity of bacterial detection in cerebrospinal fluid by fluorescent staining on low-fluorescence membrane filters.
AuthorsDurtschi JD, Erali M, Bromley LK, Herrmann MG, Petti CA, Smith RE, Voelkerding KV
JournalJ Med Microbiol
PubMed ID16091435
A membrane-filter-based, fluorescent Gram stain method for bacterial detection in cerebrospinal fluid samples was developed and evaluated as a rapid, sensitive alternative to standard Gram stain protocols. A recently developed, modified version of the aluminium oxide membrane Anopore with low-fluorescence optical properties showed superior performance in this application. Other aspects ... More
Nucleic acid stains as indicators of Cryptosporidium parvum oocyst viability.
AuthorsBelosevic M, Guy RA, Taghi-Kilani R, Neumann NF, Gyürék LL, Liyanage LR, Millard PJ, Finch GR
JournalInt J Parasitol
PubMed ID9279581
We developed nucleic acid dye staining methodology for untreated, heat-treated and chemically inactivated C. parvum oocysts. The nucleic acid staining was compared to in vitro excystation and animal infectivity using split samples of oocysts. Among the nucleic acid stains tested, SYTO-9, hexidium and SYTO-59 stained the oocysts consistently, and the ... More
Electrophoretic behavior of individual nuclear species as determined by capillary electrophoresis with laser-induced fluorescence detection.
AuthorsGunasekera N, Musier-Forsyth K, Arriaga E
JournalElectrophoresis
PubMed ID12210266
We determined the feasibility of using capillary electrophoresis with postcolumn laser-induced fluorescence (CE-LIF) detection to characterize electrophoretic properties of isolated cell nuclei and impurities present in nuclear fractions. These fractions were isolated from NS-1 mouse hybridoma cells, stained with hexidium iodide, a DNA intercalating dye, and analyzed by CE-LIF detection. ... More
Simultaneous fluorescent gram staining and activity assessment of activated sludge bacteria.
Wastewater treatment is one of the most important commercial biotechnological processes, and yet the component bacterial populations and their associated metabolic activities are poorly understood. The novel fluorescent dye hexidium iodide allows assessment of Gram status by differential absorption through bacterial cell walls. Differentiation between gram-positive and gram-negative wastewater bacteria ... More
Influence of trans-membrane potential and of hydrophobic interactions on dye accumulation in mitochondria of living cells. Photoaffinity labelling of mitochondrial proteins, action of potential dissipating drugs, and competitive staining.
AuthorsSchneider K, Naujok A, Zimmermann HW
JournalHistochemistry
PubMed ID7960945
The lipophilic cationic fluorescent dye azopentylmethylindocarbocyanine (APMC) specifically stains the mitochondria in living cells. The dye contains a photosensitive diazirine ring and is suitable for photoaffinity labelling of mitochondrial proteins. By a combination of photoaffinity labelling cell cultures of mouse fibroblasts (LM) with APMC, lysis of the labelled cells, subsequent ... More
Potential for broad applications of flow cytometry and fluorescence techniques in microbiological and somatic cell analyses of milk.
AuthorsGunasekera TS, Veal DA, Attfield PV
JournalInt J Food Microbiol
PubMed ID12878385
Monitoring the quality and safety of milk requires careful analysis of microbial and somatic cell loading. Our aim was to demonstrate proof of the principle that flow cytometry (FCM), coupled with fluorescence techniques for distinguishing between cell types, could potentially be employed in a wide variety of biological assays relevant ... More
Macrophage Phagocytosis Assay of Staphylococcus aureus by Flow Cytometry.