IF Protocol Hub Practical immunofluorescence guidance
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Protocols

Begin with the specimen, not a universal recipe.

The protocol pages share a common structure: planning questions, controls, staged workflow, stop points, imaging evidence, and conditions that require local validation.

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 evidence-based 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 →