Methods and devices for reading microarrays
View Patent ↗In one embodiment of the invention, a method to image a probe array is described that includes focusing on a plurality of fiducials on a surface of an array. The method utilizes obtaining the best z position of the fiducials and using a surface fitting algorithm to produce a surface fit profile. One or more surface non-flatness parameters can be adjusted to improve the flatness image of the array surface to be imaged.
1. A method for improving image flatness of a surface image of a probe array having an array surface roughness, the method comprising:
imaging a plurality of fiducials at a plurality of z positions and determining a best z position measurement for each of the plurality of fiducials at which the imaging is sharpest;
generating a surface fit profile based on the best z position measurement for each of the plurality of fiducials; and
imaging the probe array and adjusting one or more surface non-flatness parameters based on the surface fit profile to improve the image flatness of the surface image of the probe array.
2. The method of claim 1 , wherein the surface fit profile is constructed using a surface-fitting algorithm.
3. The method of claim 2 , wherein the surface-fitting algorithm comprises a least square algorithm.
4. The method of claim 2 , wherein the surface-fitting algorithm comprises a sub-plane surface fit.
5. The method of claim 2 , wherein the surface-fitting algorithm comprises a B spline surface fit.
6. The method of 1 , wherein the determining step comprises using quadratic interpolation.
7. The method of claim 1 , wherein the one or more surface non-flatness parameters comprises a probe array tilt angle.
8. The method of claim 1 , wherein the one or more surface non-flatness parameters comprise positioning of a stage on which the probe array is mounted.
9. The method of claim 1 , wherein the one or more surface non-flatness parameters comprise an optical parameter.
10. The method of claim 1 , wherein the one or more surface non-flatness parameters comprise a focal plane position.
11. The method of claim 1 , wherein the plurality of fiducials comprises 4 fiducials.
12. The method of claim 11 , wherein the plurality of fiducials comprises 5 fiducials.
13. The method of claim 12 , wherein the plurality of fiducials comprises 9 fiducials.
14. The method of claim 13 , wherein the plurality of fiducials comprises 12 fiducials.
15. The method of claim 14 , wherein the plurality of fiducials comprises 15 fiducials.