Method for determining whether magnetic bead in microwell

Abstract

A method for determining whether magnetic beads are present in a micropore includes the following steps: providing a micropore array wafer, wherein the micropore array wafer has a micropore array region and a plurality of micropores arranged in an array in the micropore array region; injecting a first liquid containing a plurality of magnetic beads into the micropore array wafer to disperse the plurality of magnetic beads within the plurality of micropores; capturing a bright field image of the micropore array region, wherein the bright field image includes a plurality of first regions and a plurality of second regions, the plurality of first regions corresponding to the plurality of micropores, and the plurality of second regions corresponding to the regions between the plurality of micropores; calculating the standard deviation of a plurality of signal intensity values ​​of a plurality of pixels in each first region; and when the standard deviation value corresponding to one of the plurality of first regions is greater than a first threshold, it indicates that a magnetic bead is present in the micropore corresponding to that region.

Claims

1. A method for determining whether there are magnetic beads inside a micropore, comprising: A micropore array wafer is provided, wherein the micropore array wafer has a micropore array region and a plurality of micropores arranged in an array in the micropore array region; A first liquid containing multiple magnetic beads is injected into the micro-hole array wafer to disperse the multiple magnetic beads within the multiple micro-holes; a bright-field image of the micro-hole array region is captured, wherein the bright-field image includes multiple first regions and multiple second regions, the multiple first regions corresponding to the multiple micro-holes, and the multiple second regions corresponding to the regions between the multiple micro-holes; the standard deviation values ​​of multiple signal intensity values ​​of multiple pixels in each of the multiple first regions are calculated; and when the standard deviation value corresponding to one of the multiple first regions is greater than a first threshold, it indicates that there are magnetic beads in the micro-hole corresponding to that region.

2. The method of claim 1, wherein the diameter of the plurality of micropores is 5 micrometers to 15 micrometers and the depth of the plurality of micropores is 3 micrometers to 10 micrometers.

3. The method as described in claim 1, wherein the diameter of the magnetic bead is from 1 micrometer to 10 micrometers.

4. The method as described in claim 1, wherein the material of the magnetic bead is a superparamagnetic material.

5. The method of claim 1, wherein the material of the microporous array wafer includes polycarbonate, cyclic olefin copolymer, cyclic olefin polymer or polymethyl methacrylate.

6. The method of claim 1, wherein the micropore array wafer comprises: A microporous array substrate having the microporous array region and including the plurality of micropores; A cover plate is disposed opposite to the microporous array substrate; And microchannels, disposed between the cover plate and the microporous array substrate.

7. The method of claim 6, wherein injecting the first liquid containing the plurality of magnetic beads into the micropore array wafer to disperse the plurality of magnetic beads within the plurality of micropores comprises: The first liquid containing the plurality of magnetic beads is injected into the microchannel; A magnet is positioned below the micro-hole array wafer to cause the plurality of magnetic beads to settle onto the surface of the micro-hole array substrate or into the plurality of micro-holes; And inject a second liquid into the microchannel to remove the first liquid in the microchannel, and allow the plurality of magnetic beads settled on the surface to enter the plurality of micropores.

8. The method of claim 1, wherein the micropore array region has a plurality of micropore units, one of the plurality of micropore units being disposed corresponding to one of the plurality of micropores, and the side length of the plurality of micropore units being 10 micrometers to 30 micrometers.

9. A method for determining whether there are magnetic beads inside a micropore, comprising: A micropore array wafer is provided, wherein the micropore array wafer has a micropore array region and a plurality of micropores arranged in an array in the micropore array region; A first liquid containing multiple magnetic beads is injected into the micro-hole array wafer to disperse the multiple magnetic beads within the multiple micro-holes; a bright-field image of the micro-hole array region is captured, wherein the bright-field image includes multiple first regions and multiple second regions, the multiple first regions corresponding to the multiple micro-holes, and the multiple second regions corresponding to the regions between the multiple micro-holes; a standard deviation and a first average value of multiple signal intensity values ​​of multiple pixels in each of the multiple first regions are calculated; and when the standard deviation value corresponding to one of the multiple first regions is greater than a first threshold and the first average value is greater than a second threshold, it indicates that there are magnetic beads in the micro-hole corresponding to that region.

10. A method for determining whether there are magnetic beads inside a micropore, comprising: A micropore array wafer is provided, wherein the micropore array wafer has a micropore array region and a plurality of micropores arranged in an array in the micropore array region; A first liquid containing multiple magnetic beads is injected into the micro-hole array wafer to disperse the multiple magnetic beads within the multiple micro-holes; a bright-field image of the micro-hole array region is captured, wherein the bright-field image includes multiple first regions and multiple second regions, the multiple first regions corresponding to the multiple micro-holes, and the multiple second regions corresponding to the regions between the multiple micro-holes; a standard deviation and a first average value of multiple signal intensity values ​​of multiple pixels in each of the multiple first regions are calculated, and a second average value of multiple signal intensity values ​​of multiple pixels in each of the multiple second regions is calculated; and when the standard deviation value corresponding to one of the multiple first regions is greater than a first threshold, and the difference between the first average value corresponding to that region and the second average value corresponding to the adjacent second region is greater than a third threshold, it indicates that there are magnetic beads in the micro-hole corresponding to that region.

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