Photoinduced signal amplification through externally sensitized photofragmentation in masked photosensitizers and photoamplified fluorescence turn-off system
First Claim
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1. A method of amplified fluorescence quenching comprising:
- (a) providing a plurality of masked photosensitizers, each masked photosensitizer having a masking group bonded to a quenching photosensitizer through a releasable covalent bond which disrupts the conjugation of the quenching photosensitizer;
(b) providing a fluorophor in quenching proximity to a first masked photosensitizer;
(c) providing a reaction photosensitizer in releasing proximity to a masked photosensitizer, exciting the reaction photosensitizer with photoradiation, whereby the reaction photosensitizer releases the masking group from a masked photosensitizer, forming an unmasked photosensitizer;
(d) exciting the unmasked photosensitizer with photoradiation, whereby this excited unmasked sensitizer induces fragmentation in a masked photosensitizer, inducing release of the masking group from the first masked photosensitizer, producing a quenching photosensitizer which (1) reduces the fluorescence of the fluorophor and (2) induces the release of the masking group from another masked photosensitizer; and
(e) repeating step (d).
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Abstract
Provided is a photoamplified fluorescence turn-off assay where a masked photosensitizer is mixed with a fluorescent molecule. This mixture is brightly fluorescent because the masked photosensitizer is not capable of quenching the fluorophore. When the photosensitizer is released and amplified, the photosensitizer quenches the emission of fluorophores very efficiently.
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17 Claims
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1. A method of amplified fluorescence quenching comprising:
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(a) providing a plurality of masked photosensitizers, each masked photosensitizer having a masking group bonded to a quenching photosensitizer through a releasable covalent bond which disrupts the conjugation of the quenching photosensitizer; (b) providing a fluorophor in quenching proximity to a first masked photosensitizer; (c) providing a reaction photosensitizer in releasing proximity to a masked photosensitizer, exciting the reaction photosensitizer with photoradiation, whereby the reaction photosensitizer releases the masking group from a masked photosensitizer, forming an unmasked photosensitizer; (d) exciting the unmasked photosensitizer with photoradiation, whereby this excited unmasked sensitizer induces fragmentation in a masked photosensitizer, inducing release of the masking group from the first masked photosensitizer, producing a quenching photosensitizer which (1) reduces the fluorescence of the fluorophor and (2) induces the release of the masking group from another masked photosensitizer; and (e) repeating step (d). - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
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Specification