Patent ID: 7998717

Claim:
A method of performing an illuminated reaction, comprising: providing a substrate having an observation region and a reaction mixture disposed thereon, wherein the reaction mixture comprises a) a single molecule of a polymerase confined within the observation region, b) a plurality of second reactants comprising substrates for the polymerase, each of which comprises a fluorescent label, c) and photodamage mitigating agents which are present in a concentration sufficient to reduce photodamage to the polymerase comprising more than one different triplet state quencher wherein the photodamage mitigating agents reduce an amount of photodamage to the polymerase during the interaction between the single molecule of the polymerase with one of the second reactants under excitation illumination of the second reactants that would occur in the absence of the photodamage mitigating agents; and illuminating the reaction mixture on the substrate having an observation region with the excitation illumination of the second reactants.