Screening Method and Screening Apparatus Using Micro-Chamber Array
A method and apparatus for screening a microorganism or protein having a useful function by using a micro-chamber array. Using the screening method and screening apparatus in accordance with the present invention, a target protein having an enzymatic activity or a microorganism producing such a protein is screened from a sample serving as a screening object by using a base plate having a plurality of micro-chambers arranged in the form of an array. With this method and apparatus, a protein having an enzymatic activity or a microorganism producing such a protein that is present in a micro-chamber is detected by detecting a fluorescence produced by an enzyme reaction of the protein having an enzymatic activity and a fluorescence substrate.
1 . A method for screening a protein or a microorganism, comprising the steps of:
adding a fluorescent substrate that produces fluorescence by hydrolysis onto a base plate having a plurality of micro-chambers arranged in the form of an array;
dripping a sample comprising a protein or a microorganism on the base plate; and
detecting a protein having an enzymatic activity or a microorganism producing this protein that is present inside the micro-chambers by detecting fluorescence emitted due to an enzyme reaction of the protein or the microorganism and the fluorescent substrate.
2 . The method for screening according to claim 1 , wherein
the protein is a protein expressed in a surface layer of a cell membrane of a microorganism; and
in the step of detecting, the microorganism that expresses the protein having an enzymatic activity in a surface layer of a cell membrane is detected by detecting a fluorescing micro-chamber from among the plurality of micro-chambers.
3 . The method for screening according to claim 1 , wherein an inner diameter of the micro-chamber is 5 to 500 μm.
4 . The method for screening according to claim 1 , further comprising a step of taking out contents of the micro-chamber where the protein is contained after the step of detecting.
5 . A screening apparatus for screening a protein having an enzymatic activity or microorganisms producing the protein from a sample, the screening apparatus comprising:
a base plate having a plurality of micro-chambers arranged in the form of an array; and
a fluorescence detector that detects a fluorescence produced on the base plate, wherein
the fluorescence detector detects a protein having an enzymatic activity or a microorganism producing this protein that is present inside the micro-chamber by detecting a fluorescence produced by an enzyme reaction of the protein having an enzymatic activity with the fluorescent substrate.
6 . The screening apparatus according to claim 5 , wherein
the protein is a protein expressed in a surface layer of a cell membrane of a microorganism; and
the fluorescent detector detects the microorganism that expresses the protein having an enzymatic activity in a surface layer of a cell membrane by detecting a fluorescing micro-chamber from among the plurality of micro-chambers.
7 . The screening apparatus according to claim 5 , further comprising a micro-manipulator for taking out contents of the fluorescing micro-chamber.
8 . The screening apparatus according to claim 5 , wherein an inner diameter of the micro-chamber is 5 to 500 μm.