Microscopic body detection method and microscopic body detection device

Inventors

Noji, Hiroyuki • Ono, Takao • MINAGAWA, Yoshihiro

Assignees

Japan Science and Technology Agency

Abstract

There is provided a method of detecting a microscopic body stored in a plurality of receptacles formed separately from each other. The method, which is provided as a technique for enclosing a to-be-detected substance such as nucleic acid, protein, virus, and cell by means of a simple operation in droplets of an extremely small volume and enabling highly sensitive detection, includes the steps of (1) introducing a solvent into a space between a lower layer part in which the receptacles are formed and an upper layer part facing a surface of the lower layer part in which surface the receptacles are formed, wherein the solvent contains the microscopic body; (2) introducing gas into the space to form a droplet of the solvent in the receptacles, wherein the droplet contains the microscopic body; and (3) detecting the microscopic body present in the droplet optically, electrically, and/or magnetically.

Core Innovation

The invention provides a method of detecting a microscopic body stored in a plurality of receptacles formed separately from each other. The substrate includes a lower layer part in which the plurality of receptacles and sidewalls separating the respective receptacles from each other are formed, and an upper layer part facing the lower layer part. A space resides between the lower layer part and the upper layer part, and the receptacles each have a cavity and an opening and are connected to the space.

A hydrophilic solvent containing the microscopic body is introduced into the space and further introduced into each receptacle via the opening to fill up the cavity of the receptacles. Gas is then introduced into the space to substitute the hydrophilic solvent containing the microscopic body in the space by the gas, thereby disconnecting fluid connection via the space between the receptacles while leaving the hydrophilic solvent containing the microscopic body filled in the cavity of the receptacles.

Transpiration of the hydrophilic solvent that fills up the cavity of the receptacles is suppressed by a hydratable substance contained in the hydrophilic solvent, wherein the hydratable substance is a gel, a hydrophilic polymer, or an osmolyte. The microscopic body in the receptacles is detected optically, electrically, and/or magnetically, and in optically detecting embodiments the detection is based on a change in absorbance and/or fluorescence of a chromogenic substrate present in the receptacles.

Claims Coverage

The document contains four independent claims, and the core inventive features across them are the separately formed receptacles on a layered substrate, solvent loading via openings, gas substitution to disconnect fluid connection between receptacles, detection of the microscopic body, and transpiration suppression using a hydratable substance.

Across the independent claims, the core claim coverage is centered on loading a microscopic body-containing hydrophilic solvent into separately formed receptacle cavities through openings on a layered substrate, then substituting the inter-receptacle space with gas to disconnect fluid connection between receptacles while keeping the receptacle cavities filled. Detection is performed optically, electrically, and/or magnetically, including absorbance/fluorescence change of a chromogenic substrate for optical embodiments, and solvent transpiration is suppressed using a hydratable substance contained in the hydrophilic solvent.

Stated Advantages

Documented Applications