Hydrogen sulfide (H₂S) is an important reactive sulfur species that is involved in many biological functions, and H₂S imbalances have been indicated as a potential biomarker for various diseases. Different H₂S donors have been developed to deliver H₂S directly to biological systems, but few reports include donors with optical responses that allow for tracking of H₂S release. Moreover, donor systems that use the same chemistry to deliver H₂S across a palette of fluorescent responses remain lacking. Here we report five thiol-activated fluorescence turn-on COS/H₂S donors that utilize blue, yellow, orange, red, and near infrared-emitting dyes functionalized with an H₂S-releasing sulfenyl thiocarbonate scaffold. Upon treatment with thiols, each donor provides a fluorescence turn-on response (3–310-fold) and high H₂S release efficiencies (>60 %). Using combined electrode and fluorescence experiments, we directly correlate the measured H₂S release with the fluorescence response. All donors are biocompatible and release H₂S in live cell environments. In addition, we demonstrate that the NIR donor allows for imaging H₂S release in live rats via subcutaneous injection of the donor loaded into an alginate gel, which to the best of our knowledge is the first in vivo tracking of H₂S release from a fluorogenic donor in non-transparent organisms.