摘要:
A fluorescence detecting method includes the steps of collecting a first fluorescence signal of the fluorescence received by a light receiving unit when the analyte passes a position irradiated with a laser beam, collecting a second fluorescence signal of the fluorescence received by the light receiving unit in the absence of the analyte at the position irradiated with the laser beam, and adjusting a first phase difference information on the first fluorescence signal with respect to the modulation signal by using a second phase difference information on the second fluorescence signal with respect to the modulation signal to obtain a third phase difference information on the fluorescence signal of the fluorescence, and obtaining a fluorescence relaxation time constant of the fluorescence based on the third phase difference information thus obtained.
摘要:
Among donor molecules labeling protein in living cells to be measured, the rate of donor molecules binding to an acceptor molecule and occurring FRET is determined. In a plurality of previous measurement samples having different ratios of first molecule concentration to second molecule concentration, a fluorescence lifetime of the first molecule are calculated and the fluorescence lifetime minimum value of the first molecule is calculated. The samples are irradiated with a laser beam having time-modulated intensity and the fluorescence emitted by the laser-irradiated measurement samples are measured. By using the fluorescent signals thus measured, the fluorescence lifetime of the first molecule is calculated. By using the fluorescence lifetime minimum value of the first molecule and the fluorescence lifetime of the first molecule that is calculated above, the rate of the first molecules occurring FRET in the first molecules in the measurement samples is calculated.
摘要:
A fluorescence detection device for a flow site meter emits laser light intensity-modulated in accordance with a modulation signal and acquires a fluorescent signal of fluorescence emitted from a measurement object that passes through a measurement point of the laser light. The fluorescence detection device generates, separately from the modulation signal, a reference signal having a frequency different from a frequency of the modulation signal and a phase in synchronization with a phase of the modulation signal. The fluorescence detection device determines a fluorescent relaxation time of the measurement object from the fluorescent signal by using the reference signal.
摘要:
Provided are a fluorescence detecting method and a fluorescence detecting device for calculating the fluorescence relaxation time constant more accurately than in the prior art when fluorescence emitted from an analyte is detected by applying laser beam. When the analyte passes the position irradiated with laser beam modulated in intensity by a modulation signal of a predetermined frequency, a first fluorescence signal of fluorescence received by a light-receiving means is collected. A second fluorescence signal of the fluorescence received by the light-receiving means is collected in a state in which the analyte is not in the position irradiated with the laser beam after the analyte has passed the position irradiated with the laser beam. Phase difference information of a fluorescence signal of the fluorescence emitted from the analyte with respect to the modulation signal of the laser beam is obtained by using the collected first fluorescence signal and second fluorescence signal, and the fluorescence relaxation time constant of the fluorescence emitted from the analyte is obtained from the obtained phase difference information of the fluorescence signal of the fluorescence emitted from the analyte.
摘要:
Fluorescence detection device employed in a flow site meter emits laser light intensity-modulated by a modulation signal and acquires the fluorescence signal of fluorescence emitted from a measurement object passing through a measurement point of the laser light. The device generates the reference signal, separately from the modulation signal, the reference signal having a frequency different from the frequency of the modulation signal and having a phase synchronized with a phase of the modulation. The device determines fluorescence relaxation time of the measurement object from the fluorescence signal using the reference signal.
摘要:
Disclosed herein is a fluorescence detecting apparatus. The fluorescence detecting apparatus includes a light-receiving element that receives fluorescence emitted from an object to be measured irradiated with laser light modulated at a predetermined frequency and outputs a fluorescence signal at an adjusted output level; a signal processing unit that mixes the outputted fluorescence signal and a modulation signal with the frequency to generate fluorescence data including information about phase and intensity; and an analyzing device that calculates a first phase shift of the fluorescence emitted from the object to be measured with respect to the modulation signal, calculates a second phase shift by correcting the calculated first phase shift depending on conditions for adjusting the output level, and calculates a fluorescence relaxation time of the fluorescence emitted from the object to be measured using the calculated second phase shift.
摘要:
Fluorescence detection device employed in a flow site meter emits laser light intensity-modulated by a modulation signal and acquires the fluorescence signal of fluorescence emitted from a measurement object passing through a measurement point of the laser light. The device generates the reference signal, separately from the modulation signal, the reference signal having a frequency different from the frequency of the modulation signal and having a phase synchronized with a phase of the modulation. The device determines fluorescence relaxation time of the measurement object from the fluorescence signal using the reference signal.
摘要:
A fluorescence detecting device irradiates an object to be measured with a laser beam, receives fluorescence generated from the object and processes a fluorescence signal generated when receiving the fluorescence. The device includes a laser light source section outputting the laser beam for irradiating the object, a light receiving section outputting the fluorescence signal of the fluorescence generated by the irradiated object, a light source control section generates a modulation signal having a frequency in order to time-modulate an intensity of the laser beam, and a processing section that calculates a fluorescence relaxation time of the fluorescence of the object based on the fluorescence signal output from the light receiving section. From the detection values acquired by the device including the phase information on the fluorescence, the intensity of the fluorescence is calculated.
摘要:
A fluroescene detecting device irradiates an object to be measured with a laser beam, receives fluroescene generated from the object and processes a fluorescence signal generated when receiving the fluroescene. The device includes a laser light source section outputting the laser beam for irradiating the object, a light receiving section outputting the fluorescene signal of the fluorescene generated by the irradiated object, a light source control section generates a modulatioon signal having a frequency in order to time-modulate an intensity of the laser beam, and a processing section that calculates a flurescene relaxation time of the fluorescene of the object based on the fluorescene relaxation time of the fluorescene of the object based on the fluorescene relaxation time of the fluorescene of the object based on the fluorescene signal output from the light receiving section. From the detection values acquired by the device including the phase information on the fluorescene, the intensity of the fluorescene is calculated.
摘要:
Disclosed herein is a method for measuring FRET by irradiating with laser light a measurement sample. FRET is transfer of energy from a first molecule to a second molecule. The first molecule and the second molecule are included in the measurement sample in which ligands are bound to receptors. The method includes the steps of: irradiating the measurement sample with laser light; measuring fluorescence emitted by the measurement sample; calculating a fluorescence lifetime of the first molecule; calculating a binding ratio; setting a binding condition for the measurement sample; and calculating a dissociation constant. In the dissociation constant calculating step, the dissociation constant is determined by using a least-squares method to fit a function having, as variables, a total concentration of the receptor in the measurement sample and the dissociation constant to the binding ratio calculated in the binding ratio calculating step.