System for the mitigation of photodamage in analytical reactions
First Claim
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1. A system for analyzing an illuminated reaction that is susceptible to photodamage, comprising:
- a substrate surface having reagents for the reaction disposed thereon, the reagents comprising a first reactant that comprises a polymerase, a second reactant that comprises a fluorescent or fluorogenic substrate for the polymerase, and more than one different photodamage mitigating agent present in a concentration sufficient to reduce photodamage to the polymerase when the first and second reactant are subjected to an excitation illumination for the second reactant, wherein interaction of the first reactant with the second reactant under the excitation illumination causes photodamage to the polymerase in the absence of the more than one different photodamage mitigating agent,a mounting stage supporting the substrate surface and configured to receive the substrate surface;
an optical train positioned to be in optical communication with at least a first portion of the substrate surface to illuminate the first portion of the substrate surface and detect a first set of signals emanating therefrom;
a translation system operably coupled to the mounting stage or the optical train for moving one of the optical train and the substrate surface relative to the other to newly illuminate a second portion of the substrate surface and detect a second set of signals emanating therefrom.
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Abstract
Compositions, devices, systems and methods for reducing and/or preventing photodamage of one or more reactants in illuminated analytical reactions by one or more of incorporating photodamage mitigating agents within the reaction mixture and/or interrogating different observation regions of the reaction mixture for a period that is less than a photodamage threshold period.
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Citations
8 Claims
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1. A system for analyzing an illuminated reaction that is susceptible to photodamage, comprising:
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a substrate surface having reagents for the reaction disposed thereon, the reagents comprising a first reactant that comprises a polymerase, a second reactant that comprises a fluorescent or fluorogenic substrate for the polymerase, and more than one different photodamage mitigating agent present in a concentration sufficient to reduce photodamage to the polymerase when the first and second reactant are subjected to an excitation illumination for the second reactant, wherein interaction of the first reactant with the second reactant under the excitation illumination causes photodamage to the polymerase in the absence of the more than one different photodamage mitigating agent, a mounting stage supporting the substrate surface and configured to receive the substrate surface; an optical train positioned to be in optical communication with at least a first portion of the substrate surface to illuminate the first portion of the substrate surface and detect a first set of signals emanating therefrom; a translation system operably coupled to the mounting stage or the optical train for moving one of the optical train and the substrate surface relative to the other to newly illuminate a second portion of the substrate surface and detect a second set of signals emanating therefrom. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
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Specification