Table 1:
Controls.
| Control | Purpose | Ckn-anti-GFAP | Gp-anti-SYN | Rb-anti-Iba1 | Rt-anti-CD68 | Gt-anti-Ckn 647 | Gt-atnti-Gp Biotinylated | Streptavidin 490LS | Gt-anti-Rb 594 | Gt-anti-Rt 488 | |
|---|---|---|---|---|---|---|---|---|---|---|---|
| No-primaries | No-GFAP | Ensure non-specific binding and spectral cross talk are measured and/or eliminated | − | + | + | + | + | + | + | + | + |
| No-synapsin | + | − | + | + | + | + | + | + | + | ||
| No-Iba1 | + | + | − | + | + | + | + | + | + | ||
| No-CD68 | + | + | + | − | + | + | + | + | + | ||
| No-label | Measure autofluorescence and background | − | − | − | − | − | − | − | − | − | |
| Channel alignment | Measure chromatic aberration | − | + | − | − | * | + | + | * | * | |
*
Replace fluorophores targeting other host species with fluorophores targeting the host species of the primary antibody used in the channel alignment control. For above example, one would use Gt-α-Gp 647, Gt-α-Gp 594, and Gt-α-Gp 488 in lieu of the other fluorophores.
Abbreviations: GFAP, Glial Fibrillary Acidic Protein; Ckn, chicken; SYN, synapsin; Gp, guinea pig; Rb, rabbit; Rt, rat; Gt, goat.