Table 2 Final optimised sequence of multispectral immunofluorescence staining.

From: Automated quantitative high-throughput multiplex immunofluorescence pipeline to evaluate OXPHOS defects in formalin-fixed human prostate tissue

Cycle

Primary antibody

Amplification system

Fluorophore

1

NDUFB8 (Anti-Ms IgG1, 60 mins)

Anti-mouse HQ (16 mins), followed by anti-HQ HRP (16 mins)

Opal 520 (8 mins)

2

TOMM20 (Anti-Rb IgG, 30 mins)

OmniMap anti-rabbit HRP (16 mins)

Opal 570 (8 mins)

3

MTCO1 (Anti-Ms IgG2a, 60 mins)

OmniMap anti-mouse HRP (16 mins)

Opal 620 (8 mins)

4

Pan-CK (Anti-Ms IgG1, 30 mins)

OmniMap anti-mouse HRP (16 mins)

Opal 690 (8 mins)

5

Spectral DAPI (8 mins)

–

–

Back to article page