Microfluidic devices and observation methods
A microfluidic device includes a substrate having an electromagnetic wave transmission property, a lid member facing the substrate and being separated from the substrate such that a flow channel is formed between the substrate and the lid member, a light absorption layer which is placed in the flow channel and absorbs an electromagnetic wave, and a microwell array formed on the substrate and having plural microwells that are open to the flow channel to receive a target of analysis.
1. A microfluidic device, comprising:
a substrate having an electromagnetic wave transmission property;
a micropore array layer formed on the substrate and having a plurality of microwells such that the plurality of microwells is formed in a microarray area of the micropore array layer and has a plurality of openings, respectively;
a lid member facing the substrate and separated from the substrate such that the lid member covers the micropore array layer on the substrate and forms a flow channel extending between the lid member and the microarray area of the micropore array layer and directly over the plurality of openings of the plurality of microwells and that the flow channel is directly communicated to the plurality of microwells through the plurality of openings and supplies an aqueous liquid comprising a target of analysis to the plurality of microwells between the substrate and the lid member; and
a light absorption layer comprising liquid and filling the flow channel between the micropore array layer and the lid member such that the light absorption layer is configured to absorb an electromagnetic wave corresponding to an observation light of the target of analysis and shields the plurality of microwells in the micropore array layer from the electromagnetic wave from the lid member and that the liquid of the light absorption layer is not readily miscible with the aqueous liquid and covers the aqueous liquid in the plurality of microwells.
2. The microfluidic device of claim 1 , wherein the light absorption layer includes a light absorption material that absorbs light of a wavelength in accordance with the target of analysis.
3. The microfluidic device of claim 1 , wherein the micropore array layer comprises a bottom layer positioned on the substrate and a wall layer formed on the bottom layer such that the wall layer has the plurality of microwells formed in a microwell array area of the wall layer and having the plurality of openings at the flow channel, respectively.
4. The microfluidic device of claim 1 , wherein the lid member has an inlet port at one end of the flow channel and an outlet port at an opposite end of the flow channel such that the plurality of microwells is formed in the microwell array area between the inlet port and the outlet port.
5. The microfluidic device of claim 3 , wherein the liquid of the light absorption layer is an oil-based sealant.
6. The microfluidic device of claim 4 , wherein the liquid of the light absorption layer is an oil-based sealant.
7. The microfluidic device of claim 1 , wherein the light absorption layer includes a colored component dispersed in the liquid.
8. A fluorescence observation kit, comprising:
the microfluidic device of claim 1 .
9. A method of detecting a target of analysis, comprising:
supplying the aqueous liquid including the target of analysis into the flow channel such that the aqueous liquid is introduced into the microwells in the microfluidic device of claim 1 prior to forming the light absorption layer in the microfluidic device;
forming the light absorption layer of the microfluidic device on the aqueous liquid introduced into the microwells such that the aqueous liquid is sealed in the microwells by the light absorption layer;
irradiating a first electromagnetic wave onto the microwells; and
detecting a second electromagnetic wave of the observation light emitted from the microwells.
10. The method of claim 9 , wherein the detecting is conducted through an outer surface of the substrate after the irradiating of the first electromagnetic wave.
11. The method of claim 10 , wherein the detecting is conducted on an observation surface located inside the microwells, which is in contact with the light absorption layer.
12. The method of claim 10 , wherein the detecting is conducted on an observation surface located inside the microwells, and the light absorption layer is located farther from the observation surface.
13. The method of claim 11 , wherein the target of analysis is one of DNA, RNA, miRNA, mRNA, and protein.
14. The method of claim 12 , wherein the target of analysis is one of DNA, RNA, miRNA, mRNA, and protein.
15. The method of claim 11 , wherein the detecting comprises detecting a signal from an enzymatic reaction when the enzymatic reaction occurs in the aqueous liquid applied on the observation surface.
16. The method of claim 12 , wherein the detecting comprises detecting a signal from an enzymatic reaction when the enzymatic reaction occurs in the aqueous liquid applied on the observation surface.
17. The method of claim 15 , wherein the target of analysis is nucleic acid, and the enzymatic reaction is an Invader reaction.
18. The microfluidic device of claim 1 , wherein the liquid of the light absorption layer comprises a sealant and a light absorbent substance soluble in the sealant.
19. The microfluidic device of claim 1 , wherein the micropore array layer is integrally molded with the substrate.
20. The method of claim 9 , wherein the micropore array layer is integrally molded with the substrate.