Immunofluorescence (IF) is a popular antibody-based technique, used to analyse the expression and distribution of proteins in tissues and cells. One of the advantages of using IF over conventional chromogenic stains is the ease of multiplex staining by utilising the distinct emission spectra of separate fluorophores to observe multiple targets on the same slide. IF also has relatively high sensitivity with a wide dynamic range, allowing the observation of low abundance proteins alongside highly expressed proteins, as well as the ability to see protein-protein co-localisations. However, one artifact of IF is the presence of autofluorescence within certain samples, which can often mask the expression of lowly expressed targets or dim dyes. Unfortunately, this makes discerning specific staining from noise in your slide challenging.
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