Method for amplification-free nucleic acid detection on optofluidic chips
An optofluidic platform is constructed so as to comprise a planar, liquid-core integrated optical waveguides for specific detection of nucleic acids. Most preferably, the optical waveguides comprises antiresonant reflecting optical waveguide (ARROWs). A liquid solution can be prepared and introduced into the optofluidic platform for optical excitation. The resulting optical signal can be collected at the edges of the optofluidic platform and can be analyzed to determine the existence of a single and/or a specific nucleic acid.
1. A method for amplification-free detection of the presence of a specific nucleic acid sequence in a sample, comprising:
introducing a liquid solution into one of hollow-core antiresonant reflecting waveguides (ARROWs) of a planar optofluidic platform, the liquid solution comprising said specific nucleic acid sequence and a nucleic acid probe labeled with fluorophores, wherein said nucleic acid probe specifically hybridizes with said specific nucleic acid sequence in the liquid solution;
inducing electrokinetic particle movement to the liquid solution in the one of the hollow-core ARROWs;
optically exciting the fluorophores using the planar optofluidic platform, thereby producing a fluorescent signal, wherein said optically exciting the fluorophores comprises injecting light into one of solid-core ARROWs of the planar optofluidic platform such that the light injected into the one of the solid-core ARROWs of the planar optofluidic platform is guided to an optical excitation region having a sub-picoliter volume at an intersection of the one of the hollow-core ARROWs and the one of the solid-core ARROWs;
collecting the fluorescent signal using a photodetector and converting the fluorescent signal to an electrical current or a voltage; and
detecting the presence of the specific nucleic acid sequence by analyzing the electrical current or the voltage such that the presence of the specific nucleic acid sequence in the sample is detected without amplification of the specific nucleic acid sequence.
2. The method of claim 1 , further comprising:
introducing a pure buffer solution into one of the hollow-core ARROWs of the planar optofluidic platform, the pure buffer solution being free of nucleic acids; and collecting a signal from the pure buffer solution.
3. The method of claim 2 , further comprising:
comparing the fluorescent signal to the signal from the pure buffer solution.
4. The method of claim 1 further comprising creating a fluorescence correlation spectrum using the fluorescent signal.
5. The method of claim 1 , wherein the specific nucleic acid sequence is from a virus.
6. The method of claim 1 , wherein the planar optofluidic platform further comprises planar, liquid-core integrated optical waveguides.
7. The method of claim 1 , wherein the solid-core ARROWs are connected to different points of the hollow-core ARROWs and the planar optofluidic platform further comprises means for introducing liquid solutions into the hollow-core ARROWs, means for providing an excitation light to the hollow-core ARROWs, and means for collecting a fluorescent signal.
8. The method of claim 1 , wherein the solid-core ARROWs are configured to connect to different points of the hollow-core ARROWs, the hollow-core ARROWs and the solid-core ARROWs are configured to provide separate access points for the liquid solution and the light injected into the one of the hollow-core ARROWs and to define optical excitation areas in the sub-picoliter volume.
9. The method of claim 1 , wherein the solid-core ARROWs are configured to connect to different points of the hollow-core ARROWs and the planar optofluidic platform further comprises reservoirs at ends of the hollow-core ARROWs configured to at least introduce liquid solutions into the hollow-core ARROWs, a path for an excitation light to enter the hollow-core ARROWs through solid-core ARROWs orthogonally intersecting the hollow-core ARROWs, and a path for collecting a fluorescent signal perpendicularly in the plane of the planar optofluidic platform and for guiding the fluorescent signal to edges of the planar optofluidic platform for detection.
10. The method of claim 1 , wherein said introducing the liquid solution is carried out using fluidic reservoirs contained in the planar optofluidic platform.
11. The method of claim 1 , wherein said optically exciting the fluorophores is carried out using an excitation light that enters the planar optofluidic platform.
12. The method of claim 1 , wherein the planar optofluidic platform is an optofluidic chip and the fluorescent signal is collected perpendicularly in the chip plane of the optofluidic chip and guided to the chip edges of the optofluidic chip for detection.
13. The method of claim 1 , further comprising creating a plot of the fluorescent signal.
14. The method of claim 1 , wherein said introducing the liquid solution comprises introducing the liquid solution as a stream of the liquid solution and moving the stream of the liquid solution past an optical excitation point contained in the planar optofluidic platform.
15. The method of claim 1 , wherein said light is a laser light.
16. The method of claim 1 , wherein said converting the fluorescent signal is carried out using a photodiode.
17. The method of claim 1 , wherein said converting the fluorescent signal is carried out using an avalanche photodiode.