Multi-color immunofluorescence microscopy in cultured cells

Cultured human amniotic fluid cells characterized with antibodies against intermediate filaments in indirect immunofluorescence microscopy.

Cells cultured from second trimester human amniotic fluid were characterized in indirect immunofluorescence (IIF) microscopy using specific antibodies against the subunit proteins of different types of cytoskeletal intermediate filaments. Most of the amniotic fluid cell cultures contained only epithelial cells as indicated by the positive keratin-fluorescence in IIF. Five distinct types of kera...

Redistribution and differential extraction of soluble proteins in permeabilized cultured cells. Implications for immunofluorescence microscopy.

Immunofluorescence microscopy is widely used to characterize the cellular distribution of both soluble and structural proteins. Control experiments generally address only the specificity of the antibodies used. The permeabilization/fixation conditions used to prepare cells for antibody application are assumed to preserve faithfully the in vivo distributions of the protein(s) being examined. We ...

Two-color STED microscopy in living cells

Diffraction-unlimited resolution provided by Stimulated Emission Depletion (STED) microscopy allows for imaging cellular processes in living cells that are not visible by conventional microscopy. However, it has so far not been possible to study dynamic nanoscale interactions because multicolor live cell STED microscopy has yet to be demonstrated and suitable labeling technologies and protocols...

Characterization of human mesenchymal stem cells by six - color immunofluorescence

Estimating Microtubule Distributions from 2D Immunofluorescence Microscopy Images Reveals Differences among Human Cultured Cell Lines

Microtubules are filamentous structures that are involved in several important cellular processes, including cell division, cellular structure and mechanics, and intracellular transportation. Little is known about potential differences in microtubule distributions within and across cell lines. Here we describe a method to estimate information pertaining to 3D microtubule distributions from 2D f...