BACKGROUND The objective of this study was to develop a method to simultaneously examine phenotype, proliferation, apoptosis, and death of antigen-stimulated porcine lymphocytes. METHODS Peripheral blood mononuclear cells (PBMCs) were isolated from pigs vaccinated with a Brachyspira hyodysenteriae bacterin. RESULTS Once isolated, PBMCs were stained with the fluorescent membrane intercalatin...

Multicellular marine plants were collected from their natural habitats and the quantum efficiency of their photosynthesis was determined in the laboratory in five narrow wave length bands in the visible spectrum. The results along with estimates of the relative absorption by the various plastid pigments show a fairly uniform efficiency of 0.08 molecules O(2) per absorbed quantum for (a) chlorop...

We developed the chemistry, instrumentation, and software technologies needed to measure, simultaneously and independently, eight different fluorescent molecules on individual cells. Conjugation of these fluorochromes to monoclonal antibodies is straightforward; all immunofluorescence staining is accomplished with direct stains only. We built a hybrid flow cytometer with eight fluorescence dete...

The proximity ligation assay (PLA) has previously been used for the sensitive and specific detection of single proteins. In order to adapt PLA methods for the detection of cell surfaces, we have generated multivalent peptide-oligonucleotide-phycoerythrin conjugates ('burrs') that can bind adjacent to one another on a cell surface and be ligated together to form unique amplicons. Real-time PCR d...

We present experiments in which single proteins were imaged and tracked within mammalian cells. Single proteins of R-phycoerythrin (RPE) were imaged by epifluorescence microscopy in the nucleoplasm and cytoplasm at 71 frames/s. We acquired two-dimensional trajectories of proteins (corresponding to the projection of three-dimensional trajectories onto the plane of focus) for an average of 17 fra...

Laser-induced fluorescence emission (L.I.F.E.) images were obtained in situ following 532 nm excitation of cryoconite assemblages in the ice covers of annual and perennially frozen Antarctic lakes during the 2008 Tawani International Expedition to Schirmacher Oasis and Lake Untersee in Dronning Maud Land, Antarctica. Laser targeting of a single millimeter-scale cryoconite results in multiple ne...