IF Protocol Hub Practical immunofluorescence guidance
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Protocols

Choose the protocol by specimen type.

Use the cultured-cell, frozen-section, FFPE, or multiplex protocol below. Each page covers sample preparation, staining, controls, imaging, and troubleshooting for that application.

Four-panel selector comparing cultured cells, frozen tissue, FFPE tissue, and multiplex immunofluorescence.
Sample preparation determines which fixation, access, retrieval, adhesion, and imaging decisions matter most.
Cells

Cultured-cell immunofluorescence

For adherent cells grown on coverslips or chamber slides.

Target location · cell health · fixation Open guide → Cryosections

Frozen-section immunofluorescence

For OCT-embedded sections where adhesion and handling are critical.

Section integrity · fixation · autofluorescence Open guide → FFPE

Paraffin-section immunofluorescence

For formalin-fixed paraffin-embedded tissue requiring deparaffinization and antigen retrieval.

Retrieval · tissue background · morphology Open guide → Multiplex

Multiplex immunofluorescence

For simultaneous or sequential detection of multiple antigens.

Antibody species · fluorophores · single-color controls Open guide →