Device and methods for quantifying analytes introduction
Devices and methods for measuring the quantity of multiple analytes in a sample can include a device designed such that each of the analyte sensing elements is configured to measure the quantity of a predetermined analyte and machine executable instructions configured to select the proper analyte sensing element corresponding to the analyte to be measured.
1. A method of analyzing a sample to detect multiple analytes within a single reaction volume, the method comprising:
a) providing a sample having or suspected as having a first analyte, a second analyte, or both;
b) contacting the sample with a first reporter molecule configured to associate with the first analyte and a second reporter molecule configured to associate with the second analyte, the first and second reporter molecules having different emission spectra;
c) exposing the reaction volume to excitation light within a first wavelength range to cause the first reporter molecule to emit light;
d) measuring emission light emanating from the first reporter molecule to thereby detect the presence or absence of the first analyte;
e) exposing the reaction volume to excitation light within a second wavelength range to cause the second reporter molecule to emit light; and
f) measuring emission light emanating from the second reporter molecule to thereby detect the presence or absence of the second analyte,
wherein the excitation light within a first wavelength range and the excitation light within a second wavelength range are provided by different energy sources.
2. The method of claim 1 , wherein the first analyte, second analyte, or both are independently selected from the group consisting of drugs, antigens, haptens, antibodies, proteins, peptides, amino acids, hormones, steroids, cancer cell markers, tissue cells, viruses, vitamins, nucleic acids, metal ions, enzymes, lipids, radioactive isotopes, viruses, bacteria, pathogens, chemical contaminants, and pesticides.
3. The method of claim 1 , wherein the first analyte, second analyte, or both are independently selected from the group consisting of DNA, RNA, protein, eukaryotic or prokaryotic cells, carbohydrates, lipids, and metal ions.
4. The method of claim 1 , further comprising quantifying the first analyte, the second analyte, or both.
5. The method of claim 4 , wherein quantifying the first analyte, the second analyte, or both comprises comparing emission intensity measured from the sample to an analyte-specific standard curve to calculate a concentration and/or quantity of the respective analyte in the sample.
6. The method of claim 5 , further comprising generating the analyte-specific standard curve by measuring two or more standard samples of the corresponding analyte.
7. The method of claim 6 , wherein at least one of the standard samples is a sample that omits the corresponding analyte, and another standard sample is a sample that includes a known concentration of the corresponding analyte.
8. The method of claim 1 , wherein the step of exposing the reaction volume to excitation light within the first wavelength range and the step of exposing the reaction volume to excitation light within the second wavelength range are performed simultaneously.
9. The method of claim 1 , wherein the step of measuring emission light emanating from the first reporter molecule and the step of measuring emission light emanating from the second reporter molecule are performed simultaneously.
10. The method of claim 1 , further comprising separating unbound reporter molecules from the reaction volume prior to exposing the reaction volume to excitation light within the first or second wavelength range.
11. The method of claim 1 , wherein exposing the reaction volume to excitation light within the first wavelength range comprises filtering the excitation light using a first excitation filter.
12. The method of claim 11 , wherein exposing the reaction volume to excitation light within the second wavelength range comprises filtering the excitation light using a second excitation filter, wherein the first and second excitation filters are different from one another.
13. The method of claim 1 , wherein measuring emission light emanating from the first reporter molecule comprises filtering the emission light using a first emission filter.
14. The method of claim 13 , wherein measuring emission light emanating from the second reporter molecule comprises filtering the emission light using a second emission filter, wherein the first and second emission filters are different from one another.
15. The method of claim 1 , wherein the reaction volume is contained within a microcentrifuge tube, a petri dish, a cuvette, or a well plate.
16. The method of claim 1 , wherein the method is performed using an instrument comprising:
a receptacle for holding a container in which the reaction volume is disposed;
a photodetector; and
first and second analyte sensing elements (ASEs), each ASE including an energy source,
an excitation filter, and an emission filter,
wherein the first ASE functions to expose the reaction volume to excitation light within the first wavelength range and functions to filter the emission light emanating from the first reporter molecule toward the photodetector, and
wherein the second ASE functions to expose the reaction volume to excitation light within the second wavelength range and functions to filter the emission light emanating from the second reporter molecule toward the photodetector.
17. The method of claim 16 , the instrument further comprising a computer processing unit with machine executable instructions configured to automatically select the ASE corresponding to a selected analyte to be detected.