Detection method and detection apparatus of substance in biological sample
View Patent ↗A high accuracy method for detecting a biological-related substance is provided by which an abnormal state, such as the adhesion of dust, the reduction of a sample solution, or the like can be judged. Means for detecting a light emitted by the portion for light detection after dividing the light into at least a plurality of wavelength zones is provided. One of the plurality of wavelength zones includes substantially the same wavelength zone as that of the component of the excitation light. The light intensity of the component of the excitation light is detected and it is compared with a predetermined intensity (threshold). A highly accurate fluorescence measurement can be realized, by which the abnormality of a sample can be judged.
1. A method for detecting a target biological-related substance, comprising the steps of:
(a) labeling a biological-related substance using a fluorophore,
(b) capturing the biological-related substance in a portion for light detection,
(c) irradiating an excitation light for exciting the fluorophore onto the portion for light detection,
(d) detecting a light emitted by the portion for light detection after dividing the light into a plurality of wavelength zones, wherein one of the plurality of wavelength zones is substantially the same as that of a wavelength component of the excitation light,
(e) simultaneously monitoring an intensity of the component of the excitation light wavelength and the intensity of the light emitted by the fluorophore,
(f) comparing the light intensity of the wavelength zone that is substantially the same as that of the component of the excitation light with a predetermined intensity range obtained from a normal sample,
(g) quantifying the target biological-related substance, and,
(h) in the case where the light intensity of the wavelength zone that is substantially the same as that of the component of the excitation light exceeds the predetermined intensity range, judging that the light intensity emitted by the fluorophore in the portion for light detection is not appropriate,
(i) in the case where the light intensity of the wavelength zone that is substantially the same as that of the component of the excitation light does not exceed the predetermined intensity range, correcting the light intensity emitted by the fluorophore using the light intensity of the wavelength zone that is substantially the same as that of the component of the excitation light.
2. The method for detecting a biological-related substance according to claim 1 , wherein in the case where the light intensity of the wavelength zone that is substantially the same as that of the component of the excitation light exceeds the predetermined intensity range, judging and displaying in real time that the fluorescence measurement in the portion for light detection is not appropriate.
3. The method for detecting a biological-related substance according to claim 1 , wherein the portion for light detection comprises a region for reaction formed on a substantially planar substrate surface.
4. The method for detecting a biological-related substance according to claim 1 , wherein a light detection is conducted in the portion for light detection after an amplification reaction is conducted or along with the amplification reaction.
5. The method for detecting a biological-related substance according to claim 1 , wherein a biological sample is carried on any one of a DNA chip, a DNA microarray, or a protein chip.
6. The method for detecting a biological-related substance according to claim 1 , wherein the biological sample is carried on a well of a microplate.
7. The method for detecting a biological-related substance according to claim 1 , wherein the biological-related substance is labeled with a plurality of types of fluorophores.
8. The method for detecting a biological-related substance according to claim 1 , wherein the state of a sample solution is judged by detecting the scattered of light from the sample solution.
9. A method for detecting a biological-related substance, comprising:
(a) labeling a biological-related substance or such a substance that is substantially the same as the biological-related substance using a fluorophore,
(b) adding a dye having a main absorption band in the wavelength zone which is different from that of the fluorophore,
(c) capturing the biological-related substance in a portion for light detection;
(d) irradiating an excitation light for exciting the fluorophore onto the portion for light detection, and another light to detect the absorption level of the dye,
(e) detecting a light emitted by the portion for light detection after dividing the light into a plurality of wavelength zones, wherein one of the plurality of wavelength zones is substantially the same as that of a wavelength component of the another light,
(f) simultaneously monitoring an intensity of the component of the another light wavelength and the intensity of the light emitted by the fluorophore,
(g) comparing the light intensity of the wavelength zone that is substantially the same as that of the component of the another light with a predetermined intensity range obtained from a normal sample;
(h) quantifying the biological-related substance or such a substance that is substantially the same as the biological-related substance; and,
(i) in the case where the light intensity of the wavelength zone that is substantially the same as that of the component of the another light exceeds the predetermined intensity range, judging that the light intensity emitted by the fluorophore in the portion for light detection is not appropriate,
(j) in the case where the light intensity of the wavelength zone that is substantially the same as that of the component of the another light does not exceed the predetermined intensity range, correcting the light intensity emitted by the fluorescence using the light intensity of the wavelength zone that is substantially the same as that of the component of the another light.
10. The method for detecting a biological-related substance according to claim 9 , wherein the portion for light detection comprises a region for reaction formed on a substantially planar substrate surface.
11. The method for detecting a biological-related substance according to claim 9 , wherein a light detection is conducted in the portion for light detection after an amplification reaction is conducted or along with the amplification reaction.
12. The method for detecting a biological-related substance according to claim 9 , wherein a biological sample is carried on any one of a DNA chip, a DNA microarray, or a protein chip.
13. The method for detecting a biological-related substance according to claim 9 , wherein the biological sample is carried on a well of a microplate.
14. The method for detecting a biological-related substance according to claim 9 , wherein the state of a sample solution is judged by detecting the transmitted light from the sample solution.
15. The method for detecting a biological-related substance according to claim 9 , wherein a dye solution having no effect on fluorescence measurement is added to a sample solution.
16. The method for detecting a biological-related substance according to claim 9 , wherein the biological-related substance is labeled with a plurality of types of fluorophores.