Nano-fEM: Protein Localization Using Photo-activated Localization Microscopy and Electron Microscopy

Nano-fEM: protein localization using photo-activated localization microscopy and electron microscopy.

Mapping the distribution of proteins is essential for understanding the function of proteins in a cell. Fluorescence microscopy is extensively used for protein localization, but subcellular context is often absent in fluorescence images. Immuno-electron microscopy, on the other hand, can localize proteins, but the technique is limited by a lack of compatible antibodies, poor preservation of mor...

LOCALIZATION OF REOVIRUS CELL ATTACHMENT PROTEIN ?l ON THE SURFACE OF THE REOVIRION USING IMMUNOFERRITIN ELECTRON MICROSCOPY

Purified reovirus type 3 (strain Dearing) was treated with monoclonal anti-?l antibody conjugated to ferritin and examined in the electron microscope. Virion associated ferritin molecules corresponding to locations of the ?l protein were observed. Electron microsocpy of thin sections of these preparations revealed that ferritin conjugates were localized at the vertices of the viral icosahe...

A study of SeqA subcellular localization in Escherichia coli using photo-activated localization microscopy.

Escherichia coli (E. coli) cells replicate their genome once per cell cycle to pass on genetic information to the daughter cells. The SeqA protein binds the origin of replication, oriC, after DNA replication initiation and sequesters it from new initiations in order to prevent overinitiation. Conventional fluorescence microscopy studies of SeqA localization in bacterial cells have shown that th...

Quantitative Photo Activated Localization Microscopy: Unraveling the Effects of Photoblinking

In this work we discuss how to use photophysical information for improved quantitative measurements using Photo Activated Localization Microscopy (PALM) imaging. We introduce a method that reliably estimates the number of photoblinking molecules present in a biological sample and gives a robust way to quantify proteins at the single-cell level from PALM images. We apply this method to determine...

Sono-activated ultrasound localization microscopy