Spatial quantitative analysis of fluorescently labeled nuclear structures: Problems, methods, pitfalls

Hyperspectral imaging microscopy for identification and quantitative analysis of fluorescently-labeled cells in highly autofluorescent tissue.

Standard fluorescence microscopy approaches rely on measurements at single excitation and emission bands to identify specific fluorophores and the setting of thresholds to quantify fluorophore intensity. This is often insufficient to reliably resolve and quantify fluorescent labels in tissues due to high autofluorescence. Here we describe the use of hyperspectral analysis techniques to resolve ...

Quantitative single cell monitoring of protein synthesis at subcellular resolution using fluorescently labeled tRNA

We have developed a novel technique of using fluorescent tRNA for translation monitoring (FtTM). FtTM enables the identification and monitoring of active protein synthesis sites within live cells at submicron resolution through quantitative microscopy of transfected bulk uncharged tRNA, fluorescently labeled in the D-loop (fl-tRNA). The localization of fl-tRNA to active translation sites was co...

Methods for Quantitative Analysis of the Spatial Structure of Dendrites

Quantitative Studies on Enzymes in Structures in Striated Muscles by Labeled Inhibitor Methods

Di-isopropylfluorophosphate (DFP) labeled with phosphorus-32 was applied to fragments of the diaphragm and sternomastoid muscles of the mouse, in conditions in which it saturated all available sites at the motor endplates. After adequate washing and exchange with unlabeled DFP, single endplates were obtained by microdissection and their radioactivity was found by beta track radioautography. The...

Three Different Methods for Approximate Analysis of Bar Structures

In this paper, modified solutions were compared through utilizing three different approximate methods for bar structures. The modifications considered various changes in the initial design. To authors' best of knowledge, the studies have carried out on this matter so far are not broad enough and have considerred the simeltaneous variations of size, geometry and topology on the bar structures. I...