Cell analysis in multi-through-hole testing plate
A spatially fixed library of mutated cells and a method for creating such a library. The library is created by creating a set of cells having a genetic diversity, causing the cells to multiply and express phenotypes, preparing a solution containing the cells diluted in a medium, populating a plurality of through-holes of a testing plate with the solution such that surface tension holds the solution in respective through-holes, and analyzing the phenotypes of the cells in the solution on a hole-by-hole basis. Cells expressing specified phenotypes may then be retrieved.
1 . A method for creating a spatially fixed library of mutated cells, the method comprising:
a. creating a set of cells characterized by genetic diversity;
b. causing the cells to multiply and express phenotypes;
c. preparing a solution containing the cells diluted in a medium;
d. populating a plurality of through-holes of a testing plate with the solution such that surface tension holds the solution in respective through-holes; and
e. analyzing the phenotypes of the cells in the solution on a hole-by-hole basis.
2 . The method for creating a spatially fixed library of mutated cells in accordance with claim 1 , wherein the step of creating a set of cells characterized by genetic diversity includes exposing the cells to a mutagen.
3 . The method of claim 1 , wherein the step of populating the plurality of through-holes is performed prior to causing the cells to multiply and express phenotypes.
4 . The method for creating a spatially fixed library of mutated cells in accordance with claim 1 , wherein the step of introducing the set of cells in a medium into an array of through-holes includes delivering multiple cells to each of the through-holes of the plurality of through-holes.
5 . The method for creating a spatially fixed library of mutated cells in accordance with claim 2 , wherein the step of exposing the cells to a mutagen includes using at least one of ultraviolet and chemical mutagenesis.
6 . The method for creating a spatially fixed library of mutated cells in accordance with claim 1 , wherein the step of causing the cells to multiply and express phenotypes includes incubating the cells.
7 . The method for creating a spatially fixed library of mutated cells in accordance with claim 1 , wherein the step of analyzing the phenotypes includes optically analyzing contents of the through-holes.
8 . The method for creating a spatially fixed library of mutated cells in accordance with claim 1 , wherein the step of analyzing the phenotypes includes measuring fluorogenic groups liberated by hydrolysis of a substrate.
9 . The method for creating a spatially fixed library of mutated cells in accordance with claim 1 , wherein the step of analyzing the phenotypes includes measuring chromogenic groups liberated by hydrolysis of a substrate.
10 . The method for creating a spatially fixed library of mutated cells in accordance with claim 1 , wherein the step of analyzing the phenotypes includes optically imaging a plurality of through-holes of the testing plate.
11 . The method for creating a spatially fixed library of mutated cells in accordance with claim 1 , further comprising retrieving genetic determinants expressing a specified phenotype.
12 . A method in accordance with claim 11 , wherein the step of retrieving includes robotic extraction of contents of an identified through-hole.
13 . A method in accordance with claim 11 , wherein the step of retrieving includes non-robotic extraction of contents of an identified through-hole.
14 . A library of genetic diversity comprising a set of mutated cells suspended in a medium, the medium retained by surface tension in a plurality of through-holes of a test plate, the cells induced to express phenotypes.
15 . The library of genetic diversity of claim 14 , wherein the medium additionally contains an analyte that produces a detectable characteristic.
16 . The library of genetic diversity of claim 15 , wherein said detectable characteristic is produced in an absorbance transcription assay.
17 . The library of genetic diversity of claim 15 , wherein said detectable characteristic is produced in a fluorescent transcription assay.
18 . The library of genetic diversity of claim 15 , wherein said detectable characteristic is produced in a fluorescent secreted enzyme assay.
19 . The library of genetic diversity of claim 15 , wherein said detectable characteristic is produced in a microorganism screening assay.