Measurement of monomer-oligomer distributions via fluorescence moment image analysis.
AuthorsSergeev M, Costantino S, Wiseman PW
JournalBiophys J
PubMed ID16935950
We present higher-order moment analysis of fluorescence intensity fluctuations from individual laser scanning microscopy images applied to study monomer-oligomer distributions. We demonstrate that the number densities and brightness ratios of a mixed population of monomers and oligomers can be determined by analyzing higher-order moments of the fluorescence intensity fluctuations from ... More
Analysis of several fluorescent detector molecules for protein microarray use.
AuthorsWiese R
JournalLuminescence
PubMed ID12536376
The utility of several streptavidin-linked fluorescent detector molecules was evaluated on two protein microarray platforms. Tested detector molecules included: Alexa Fluor 546; R-phycoerythrin (RPE), orange fluospheres; Cy3-containing liposomes (Large Unilamellar Vesicles, LUV) labelled with Cy3; and an RPE-antibody complex. The two array architectures tested consisted of an array of murine ... More
In situ fluorescent protein imaging with metal film-enhanced total internal reflection microscopy.
AuthorsBurghardt TP, Charlesworth JE, Halstead MF, Tarara JE, Ajtai K
JournalBiophys J
PubMed ID16565065
Fluorescence detection of single molecules provides a means to investigate protein dynamics minus ambiguities introduced by ensemble averages of unsynchronized protein movement or of protein movement mimicking a local symmetry. For proteins in a biological assembly, taking advantage of the single molecule approach could require single protein isolation from within ... More
In situ single-molecule imaging with attoliter detection using objective total internal reflection confocal microscopy.
AuthorsBurghardt TP, Ajtai K, Borejdo J
JournalBiochemistry
PubMed ID16566579
Confocal microscopy is widely used for acquiring high spatial resolution tissue sample images of interesting fluorescent molecules inside cells. The fluorescent molecules are often tagged proteins participating in a biological function. The high spatial resolution of confocal microscopy compared to wide field imaging comes from an ability to optically isolate ... More