FRET MEASUREMENT DEVICE AND FRET MEASUREMENT METHOD

摘要

FRET measurement uses a FRET probe that includes a probe element X labeled with a donor fluorescent substance and a probe element Y labeled with an acceptor fluorescent substance and enables FRET to occur when the probe element X and the probe element Y approach to each other or bind together. A test sample as a measuring object in FRET measurement contains a test object about which it is unknown whether or not it has an approaching/binding property of allowing the probe element X and the probe element Y to approach to each other or bind together or a separating property of separating from each other the probe element X and the probe element Y that are in a state where they adjoin each other or bind together. A plurality of sets of a fluorescence lifetime τsample and a ratiometry Rsample obtained by this measurement are used to judge whether or not the test object has the approaching/binding property or the separating property.

主权项

1. A FRET measurement device comprising:
a conduit through which a sample flows, the sample comprising: a FRET probe that comprises a probe element X labeled with a donor fluorescent substance and a probe element Y labeled with an acceptor fluorescent substance and enables FRET to occur when the probe element X and the probe element Y approach to each other or bind together; and a test object about which it is unknown whether or not it has an approaching/binding property of allowing the probe element X and the probe element Y to approach to each other or bind together or a separating property of separating the probe element X and the probe element Y that are in a state where they adjoin each other or bind together; a light source unit configured to emit, toward the conduit, laser light whose intensity is modulated using a modulation signal; a light-receiving unit configured to receive fluorescence emitted from the FRET probe in the sample by irradiation with the intensity-modulated laser light and outputs a fluorescent signal; a fluorescence calculating unit configured to calculate, using the fluorescent signal and the modulation signal, a fluorescence lifetime τsample of donor fluorescence emitted from the donor fluorescent substance, and further to calculate, using the fluorescent signal, a ratio Rsample of fluorescence intensity of acceptor fluorescence emitted from the acceptor fluorescent substance of the FRET probe to fluorescence intensity of donor fluorescence emitted from the donor fluorescent substance so that a plurality of sets of the fluorescence lifetime τsample and the ratio Rsample are calculated; and a judgment unit configured to judge whether or not the test object has the approaching/binding property or the separating property, using the sets of the fluorescence lifetime τsample and the ratio Rsample.