Table 1: Fluorophores and cytometer optical specifications.
Cytometer specifications used in this protocol are listed for an instrument with a total of 4 lasers and 15 emission detectors. DDAOG detected at 645/660 nm is the form of the probe cleaved by SA-β-Gal1. Uncleaved DDAOG can exhibit low level fluorescence at 460/610 nm but is removed by wash steps in the protocol. Abbreviations: DDAO = 9H-(1,3-dichloro-9,9-dimethylacridin-2-one); DDAOG = DDAO-Galactoside; AF = autofluorescence; PE = phycoerythrin; SA-β-Gal = senescence-associated beta-galactosidase.
| Fluorophore | Detects | Ex/Em (nm) | Cytometer laser (nm) | Cytometer detector / bandpass filter (nm) |
|---|---|---|---|---|
| DDAOG | SA-B-Gal | 645/6601 | 640 | 670 / 30 |
| AF | Lipofuscin | < 600 | 488 | 525 / 50 |
| CV450 | Viability | 408/450 | 405 | 450 / 50 |
| PE | Antibody/surface marker | 565/578 | 561 | 582 / 15 |