Apparatus for amplification of nucleic acids
Described herein is a chip-based apparatus for amplifying nucleic acids, a cartridge housing the apparatus, and methods of using the apparatus for amplification of nucleic acids. More specifically, this invention provides integrated semiconductor chip, manufactured with standard semiconductor manufacturing process, with on-chip circuitry to perform thermal management and optical sensing necessary for amplification of nucleic acids. The apparatus and methods embodied in this invention makes it possible to build a disease diagnosis and prognosis tool that is easy to use, portable and disposable.
1. An apparatus for analyzing a biological sample comprising:
a) a semiconductor substrate comprising an integrated circuitry,
wherein the integrated circuitry on the semiconductor substrate comprises:
a junction temperature sensor; and
a first light sensor;
b) a biocompatible coating over the semiconductor substrate,
wherein the biocompatible coating is positioned to allow the biological sample to be in proximity to the integrated circuitry;
c) one or more processors;
d) memory storing one or more programs, the one or more programs configured to be executed by the one or more processors, the one or more programs including instructions for:
detecting a first light input measurement from the first light sensor at a first time prior to triggering a light signal from the biological sample;
detecting a second light input measurement from the first light sensor at a second time after triggering the light signal from the biological sample;
detecting a plurality of temperature input measurements from the junction temperature sensor,
wherein the plurality of temperature input measurements comprises:
a first temperature input measurement detected at the first time; and
a second temperature input measurement detected at the second time;
using the first light measurement to estimate noise signal; and
using the plurality of temperature input measurements to further estimate noise signal and improve noise suppression, thereby determining the light signal from the biological sample.
2. The apparatus of claim 1 , wherein the integrated circuitry on the semiconductor substrate further comprises a heating element.
3. The apparatus of claim 2 , wherein the heating element comprises a transistor.
4. The apparatus of claim 2 , wherein the heating element comprises a resistor.
5. The apparatus of claim 1 , wherein the integrated circuitry comprises complementary metal-oxide-semiconductor circuitry.
6. The apparatus of claim 1 , wherein the integrated circuitry further comprises a timer, wherein the timer is electrically connected to the one or more processors.
7. The apparatus of claim 1 , wherein the biocompatible coating is hydrophilic.
8. The apparatus of claim 1 , wherein the biocompatible coating is a light wavelength filter.
9. The apparatus of claim 8 , wherein the biocompatible coating comprises a member selected from the group consisting of zinc sulfide, Cryolite, glass, quartz, and a transparent polymer.
10. The apparatus of claim 8 , wherein the biocompatible coating comprises different portions with different light wavelength filtering characteristics.
11. The apparatus of claim 10 , wherein the biocompatible coating is a laminate placed over the semiconductor substrate.
12. The apparatus of claim 1 , wherein the biocompatible coating further comprises one or more nucleic acid amplification reagents deposited thereon.
13. The apparatus of claim 12 , wherein the one or more nucleic acid amplification reagents comprises one or more magnetic beads, and wherein the one or more magnetic beads comprises one or more nucleic acid probes covalently attached thereon.
14. The apparatus of claim 13 , further comprising a magnetic field generator positioned to immobilize the one or more magnetic beads on the biocompatible coating.
15. The apparatus of claim 12 , wherein the location of the one or more nucleic acid amplification reagents corresponds with the location of the first light sensor.
16. The apparatus of claim 1 , wherein the biocompatible coating comprises a sample partition, wherein the sample partition has a sample capacity, as measured by the amount of biological sample, of between 0.1 μl to 200 μl.
17. The apparatus of claim 16 , wherein the sample partition is aligned with the first light sensor.
18. The apparatus of claim 1 , wherein the integrated circuitry further comprises a digital communications device configured to communicate between the integrated circuitry and the one or more processors.
19. The apparatus of claim 1 , further comprising:
a communications interface;
a user interface element; and
a display.
20. The apparatus of claim 1 , wherein the apparatus further comprises a cooling device.
21. The apparatus of claim 1 , wherein the one or more programs further includes instructions for:
heating the semiconductor substrate; or
cooling the semiconductor substrate.
22. The apparatus of claim 1 , wherein the one or more programs further includes instructions for:
detecting a temperature input measurement from the junction temperature sensor;
comparing the temperature input measurement to an actual temperature measurement; and
calibrating the temperature input measurement.
23. The apparatus of claim 22 , wherein calibrating the temperature input measurement comprises applying a correction factor between the actual temperature measurement and the temperature input measurement.
24. The apparatus of claim 1 , wherein the memory comprises non-volatile memory.
25. The apparatus of claim 1 , further comprising a first light source positioned to illuminate the biological sample.
26. The apparatus of claim 25 , wherein the first light source is a laser or LED.
27. The apparatus of claim 25 , wherein the one or more programs further includes instructions for:
applying a first excitation light from the first light source to trigger the light signal from the biological sample.
28. The apparatus of claim 27 , wherein the one or more programs further includes instructions for:
not applying the first excitation light from the first light source to the biological sample at the first time.
29. The apparatus of claim 27 , further comprising a second light source positioned to illuminate the biological sample, wherein the one or more programs further includes instructions for:
applying a second excitation light from the second light source to trigger a light signal from the biological sample;
detecting a third light input measurement from the first light sensor at a third time after triggering the light signal from the biological sample; and
wherein the plurality of temperature input measurements further comprises a third temperature input measurement detected at the third time;
using the first light input measurement to estimate noise signal; and
using the plurality of temperature measurements to further estimate noise signal and improve noise suppression, thereby determining the light signal from the biological sample at the third time.
30. The apparatus of claim 29 , wherein the first excitation light is a different wavelength than the second excitation light.
31. The apparatus of claim 30 , wherein the one or more programs further includes instructions for:
not applying the first excitation light from the first light source to the biological sample at the first time; and
not applying the second excitation light from the second light source to the biological sample at the first time.
32. The apparatus of claim 31 , wherein the one or more programs further includes instructions for:
not applying the first excitation light from the first light source to the biological sample at the third time; and
not applying the second excitation light from the second light source to the biological sample at the second time.
33. The apparatus of claim 1 , wherein the light signal from the biological sample is triggered via a chemical reaction.
34. A method for amplification of a nucleic acid in a biological sample, the method comprising:
contacting the biocompatible coating of the apparatus of claim 1 with (i) the biological sample and (ii) reagents for amplification of nucleic acid in the sample, wherein said contacting forms a reaction mixture on the biocompatible coating of the apparatus;
using the apparatus to perform a nucleic acid amplification reaction; and
receiving results of the nucleic acid amplification reaction, the results including light emission data measured by the apparatus.
35. The method of claim 34 , wherein at least two distinct reaction mixtures are formed on the biocompatible coating of the apparatus.
36. The method of claim 34 , wherein said light emission data is selected from the group consisting of fluorescent light emission data and chemi-luminescent light emission data.