Methods and systems for manufacture of microarray assay systems, conducting microfluidic assays, and monitoring and scanning to obtain microfluidic assay results
A method of flowing a fluid with a tracer in a microfluidic channel of an assay device and detecting the tracer for determining the channel location or condition of the channel.
1. A method of using a tracer dye in at least one microfluidic channel of an assay device, which performs an assay having an assay protocol, comprising:
flowing a fluid with a tracer dye in the microfluidic channel;
illuminating the tracer dye with a light beam which causes the tracer dye to emit a fluorescent light signal;
detecting the tracer dye for at least one detection location on the channel using the fluorescent light signal emitted from the tracer dye; and
determining the detection location of the channel from the fluorescent light signal.
2. The method of claim 1 , further comprising:
during conduct of a first phase of the assay, under required assay conditions, monitoring a selected region, of the microfluidic channel to detect the tracer dye, and comparing the detected results with a standard of acceptable results.
3. The method of claim 1 wherein the flowing step is performed during a phase of the assay and the tracer dye being inert with respect to an assay fluid in which it is carried.
4. The method of claim 3 where in a detectable property of the tracer dye is used to confirm acceptable performance of the phase of the assay.
5. The method of claim 3 wherein the fluorescence intensity of the tracer dye is indicative of the concentration of the tracer dye in the channel and wherein the value of the concentration is used to confirm acceptable performance of the phase of an assay.
6. The method of claim 4 wherein the detectable property of the tracer is fluorescence intensity.
7. The methods of claim 1 , in which the illuminating and the detecting steps are performed collectively by an epifluorescent system having the light beam to excite fluorescence in the tracer dye within the microfluidic channel, and configured to translate the laser beam with the relation to the channel across a plurality of channels or along the length of the channel for performing at least one of detecting position, monitoring, and assay-reading purposes.
8. The method of claim 1 wherein the detecting step comprises using a region of interest (ROI) to detect only predetermined regions of the fluorescent light signal.
9. The method of claim 1 wherein the laser beam has an aspect ratio of at least 2:1.
10. The method of claim 1 in which, during the detecting step, relative movements along the length of the channel are used to index between monitoring locations, and to progressively read assay results, from at least one of immobilized detection elements in the channel.
11. The method of claim 1 in which, during the detecting step, relative movements along the length of the channel are used to determine the locations of the immobilized detection elements in the channel.
12. The method of claim 1 in which the microfluidic flow channel has a flow axis, along which a series of discrete, axially-spaced apart, transparent hollow flow elements are secured in fixed position, each hollow flow element having at least one axially-extending flow passage through its interior, the elements having interior and exterior surfaces extending in parallel in the direction of the channel axis, and end surfaces extending transversely to the axis, the surfaces of the elements exposed to liquid in the channel, and assay capture agent fixed to the interior surface of the elements for capture of an analyte in liquid flowing through the interior of the hollow flow elements, the assay device constructed to enable light to be transmitted into and out of the elements transversely to the flow axis for excitation and reading of fluorescence from captured analyte.
13. The method of claim 1 wherein the flowing step comprises flowing an assay fluid using a positive-displacement pump arranged to introduce a segment of liquid sample, and cause the sample to move back and forth with respect to a hollow flow element to produce capture of analyte only in the interior surface of the element, and to repeat this action for successive segments of liquid sample.
14. The method claim 1 in which a capture agent on the interior surface of a hollow detection element in the channel is configured to the define a code.
15. The method of claim 12 in which the assay is an ELISA and the assay capture agent comprises at least one of an antibody and an antigen.
16. The method of claim 1 , further comprising, flowing an assay fluid in the channel during a first phase of the assay protocol; and wherein the fluid with the tracer dye is the assay fluid and the tracer dye is inert to the first phase of the assay protocol.
17. The method of claim 1 , wherein the detecting step is performed at two detection locations along the channel and wherein the location of the channel is determined from the two detection locations.
18. The method of claim 2 , further comprising flowing a second assay fluid in the channel during a second phase of the assay protocol; and wherein step of flowing the fluid with the tracer dye is performed before the step of flowing the second assay fluid.
19. The method of claim 3 , wherein the second assay fluid comprises a detect reagent.
20. The method of claim 1 , further comprising, flowing an assay fluid in the channel during a detect phase of the assay protocol; and wherein the tracer dye is carried in the assay fluid and wherein the tracer dye is used to measure the results of the assay.
21. The method of claim 1 , wherein the detection step comprises adjusting a focus in a Z-plane to focus the emitted fluorescent light.
22. The method of claim 1 , wherein the detecting step further comprises detecting across a width of the channel at the detection location to identify at least one edge of the microfluidic channel.
23. The method of claim 8 , wherein the ROI used to detect the channel is different from the ROI used to detect assay results.
24. The method of claim 1 , wherein the detecting and illuminating steps collectively comprise translating the laser light beam in relation to the channel across a width of a plurality of channels at the detection location or along the length of the channel, for performing at least one of detecting position, monitoring and assay-reading.
25. The method of claim 1 , wherein the assay device is disposed in an X-Y stage, and the location of the channel is precisely determined relative to its location on the X-Y stage, without requiring the assay device be aligned on the X-Y stage with the same level of precision.
26. A method of using a tracer dye in at least one microfluidic channel of an assay device, which performs an assay having an assay protocol, comprising:
flowing a tracer dye in the microfluidic channel;
illuminating the tracer dye with a light beam which causes the tracer dye to emit a fluorescent light signal;
determining at least one detection location on the channel using the fluorescent light signal emitted from the tracer dye; and
wherein the assay device is disposed in an X-Y stage, and the location of the channel is precisely determined relative to its location on the X-Y stage, without requiring the assay device be aligned on the X-Y stage with the same level of precision.
27. A method of using a tracer dye in at least one microfluidic channel of an assay device, which performs an assay having an assay protocol, comprising:
flowing a tracer dye in the microfluidic channel;
illuminating the tracer dye with a light beam which causes the tracer dye to emit a fluorescent light signal;
determining at least one detection location on the channel using the fluorescent light signal emitted from the tracer dye;
wherein the flowing step is performed during a phase of the assay and the tracer dye is inert with respect to an assay fluid in which it is carried; and
wherein a detectable property of the tracer dye is used to confirm acceptable performance of the phase of the assay.