Paraformaldehyde (PFA) fixation is a fundamental procedure in cell biology that involves treating cells with a solution of PFA. The reagent crosslinks proteins and thus preserves the cellular structure at a particular moment in time. The purpose of fixation is to maintain cellular components such as proteins, DNA, and RNA in their native state and location. It is like an effective “freezing” of them in original place. This process allows cells to be stored for extended periods without degradation, enabling their detailed examination at a later time. PFA fixation is particularly useful for techniques such as immunofluorescence and immunohistochemistry, where it is crucial to preserve the cellular architecture and molecular arrangements.
Protocol
- put the PFA directly to the petri dish or well with cells
- incubate on RT for 5 mins
- 3x wash with PBS (or another suitable buffer)
- keep them under PBS for the following procedure
This protocol outlines the BASIC procedure. However, many procedures are often modified to suit a particular purpose.
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