AI Article Synopsis
- Immunofluorescence is a lab technique that uses antibodies tagged with fluorescent markers to identify specific structures within fixed and permeabilized cells.
- The process typically involves either directly using primary antibodies linked to fluorophores or utilizing a two-step method with primary and secondary antibodies to enhance labeling.
- The effectiveness of imaging the Golgi apparatus can vary based on the fixation and permeabilization methods used, and optimizing these conditions is crucial for accurate fluorescent detection of Golgi proteins.
Article Abstract
Immunofluorescence is a technique that uses antibodies and fluorophores to label structures inside cells. The cells are normally fixed and permeabilized, and then structures are labelled using primary antibodies directly conjugated to fluorophores, or, more commonly, first with an antibody against an antigen of interest followed by a secondary antibody conjugated to a fluorophore that binds to the primary antibody. Fluorescence can be visualized using widefield, confocal, or super-resolution microscopy. Here we focus on labelling of the Golgi apparatus and show that different fixation and permeabilization conditions can significantly affect labelling of Golgi proteins and describe how to optimize fluorescent detection of Golgi proteins.
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| http://dx.doi.org/10.1007/978-1-0716-2639-9_8 | DOI Listing |
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