METHODS AND DEVICES FOR MICRO-ISOLATION, EXTRACTION, AND/OR ANALYSIS OF MICROSCALE COMPONENTS IN AN ARRAY

摘要

Provided herein are devices and methods for the micro-isolation of biological cellular material. A micro-isolation device described can comprise a photomask that protects regions of interest against DNA-destroying illumination. The micro-isolation device can further comprise photosensitive material defining access wells following illumination and subsequent developing of the photosensitive material. The micro-isolation device can further comprise a chambered microfluidic device comprising channels providing access to wells defined in photosensitive material. The micro-isolation device can comprise a chambered microfluidic device without access wells defined in photosensitive material where valves control the flow of gases or liquids through the channels of the microfluidic device. Also included are methods for selectively isolating cellular material using the devices described herein, as are methods for biochemical analysis of individual regions of interest of cellular material using the devices described herein. Further included are methods of making masking arrays useful for the methods described herein. The micro-isolation devices can comprise a unique combination of barcodes in each microfluidics well, allowing two-dimensional mapping of genetic information.

主权项

1. A method comprising:
a) fabricating a distributor chip comprising:
a plurality of microfluidics channels, each microfluidics channel having an input port on a first surface and a plurality of output ports on a second surface opposite the first surface, each microfluidics channel, its input port and its plurality of output ports laying on a plane parallel to a subsequent or previous microfluidics channel, thereby obtaining parallel rows of the output ports on the second surface; b) fabricating a first overlay comprising parallel rows of microfluidics wells having positions corresponding to the parallel rows of the output ports; c) fabricating a receiver chip comprising parallel rows of microfluidics conduits having positions corresponding to the parallel rows of the microfluidics wells of the first overlay; d) fabricating a stopper layer permeable to air and impermeable to water; e) providing a support layer comprising parallel rows of openings corresponding to the parallel rows of the microfluidics conduits of the receiver chip; f) positioning the distributor chip on the first overlay, the first overlay on the receiver chip, the receiver chip on the stopper layer, and the stopper layer on the support layer; g) aligning the distributor chip, the first overlay, the receiver chip, the stopper layer and the support layer, by aligning the parallel rows of the output ports, the parallel rows of the microfluidics wells, the parallel rows of the microfluidics conduits, and the parallel rows of openings, thereby creating an aligned assembly; h) inserting a solution in each input port of each microfluidics channel of the plurality of microfluidics channels, each solution comprising same reagents and a different barcode for each input port; and j) removing the first overlay from the aligned assembly, thereby obtaining parallel rows of microfluidics wells, each row having the same reagents and a unique barcode in each row of the parallel rows of microfluidics wells of the first overlay.