Systems and methods for efficient collection of single cells and colonies of cells and fast generation of stable transfectants
View Patent ↗A plate manufactured to enable samples of cells, micro-organisms, proteins, DNA, biomolecules and other biological media to be positioned at specific locations or sites on the plate for the purpose of performing addressable analyses on the samples. Preferably, some or all of the sites are built from a removable material or as pallets so that a subset of the samples of interest can be readily isolated from the plate for further processing or analysis. The plate can contain structures or chemical treatments that enhance or promote the attachment and/or function of the samples, and that promote or assist in their analyses. Use of the plate advantageously enables the selection and sorting of cells based on dynamic phenomena and the rapid establishment of stable tranfectants.
1. A method of collecting a single, living cell comprising the steps of
arranging a plurality of living cells in an array, wherein each of the plurality of cells is located on a separate pallet within an array of pallets removably coupled to a plate,
selecting a single living cell from the plurality of living cells, wherein selecting a single living cell includes identifying a cell of interest based on two or more measurements of the cell separated in time, and
removing the selected living cell from the array by dislodging its pallet from the plate.
2. The method of claim 1 wherein the two or more measurements are made at intervals greater than 1 second.
3. The method of claim 1 wherein the step of selecting includes identifying a cell exhibiting a temporal characteristic of interest.
4. The method of claim 1 wherein the step of selecting includes identifying a cell exhibiting a dynamic characteristic of interest.
5. A method of collecting a single, living cell comprising the steps of
arranging a plurality of living cells in an array, wherein each of the plurality of cells is located on a separate pallet within an array of pallets removably coupled to a plate, wherein the cell volume is less than 1 nL,
selecting a single living cell from the plurality of living cells, and
removing the selected living cell from the array by dislodging its pallet from the plate.
6. The method of claim 5 wherein the cell volume is less than 100 pL.
7. A method of collecting a single, living cell comprising the steps of
identifying a single living cell in a plurality of living cells based on two or more measurements of the cell separated in time, wherein each of the plurality of cells is located on a separate pallet removably coupled to a plate, and
separating the selected living cell from the plurality of living cells by dislodging its pallet from the plate.
8. The method of claim 7 wherein the two or more measurements are made at intervals greater than 1 second.
9. The method of claim 7 wherein the step of identifying includes identifying a cell exhibiting a temporal characteristic of interest.
10. The method of claim 7 wherein the step of identifying includes identifying a cell exhibiting a dynamic characteristic of interest.
11. A method of collecting a colony of cells comprising the steps of
arranging a plurality of colonies of cells in an array, wherein each of the plurality of colonies of cells is located on a separate pallet within an array of pallets removably coupled to a plate, and wherein the cell volume of each cell in the colony is less than 1 nL in volume,
selecting a colony of cells from the plurality of colonies, and
removing the selected colony from the array by dislodging its pallet from the plate.
12. The method of claim 11 wherein the colony comprises less than 500 clonal cells.
13. The method of claim 11 wherein the colony comprises less than 100 clonal cells.
14. The method of claim 11 wherein the colony comprises less than 50 clonal cells.
15. The method of claim 11 wherein the cell volume of each cell in the colony is less than 100 pL in volume.
16. The method of claim 11 wherein the cell volume of each cell in the colony is less than 10 pL in volume.
17. A method of collecting a colony of cells comprising the steps of
arranging a plurality of colonies of cells in an array, wherein each of the plurality of colonies of cells is located on a separate pallet within an array of pallets removably coupled to a plate,
selecting a colony of cells from the plurality of colonies, wherein the step of selecting includes identifying a colony of interest based on two or more measurements of the colony separated in time, and
removing the selected colony from the array by dislodging its pallet from the plate.
18. The method of claim 17 wherein the two or more measurements are made at intervals greater than 1 second.
19. The method of claim 17 wherein the step of selecting includes identifying a colony exhibiting a temporal characteristic of interest.
20. The method of claim 17 wherein the step of selecting includes identifying a colony exhibiting a dynamic characteristic of interest.
21. A method comprising the steps of
transfecting or micromanism-mediated infecting a plurality of cells with DNA of interest, wherein each of the plurality of cells is located on a separate pallet removably coupled to a plate,
identifying a stable transfectant of a cell from the plurality of cells in less than 5 days after transfection or microrganism-mediated infection of the plurality of cells, and
collecting the stable transfectant of a cell by dislodging its pallet from the plate.
22. The method of claim 21 wherein the plurality of cells are arranged in an array.
23. The method of claim 21 wherein the cell is adherent to a surface.
24. A method comprising the steps of
transfecting or microrganism-mediated infecting a plurality of cells with DNA of interest, wherein each of the plurality of cells is located on a separate pallet removably coupled to a plate, wherein the cell volume is less than 1 nL
identifying a stable transfectant of a cell from the plurality of cells in less than 10 days after transfection or microrganism-mediated infection of the plurality of cells, and
collecting the stable transfectant of a cell by dislodging its pallet from the plate.
25. The method of claim 24 wherein the cell volume is less than 100 pL.
26. A method comprising the steps of
transfecting or microrganism-mediated infecting a plurality of cells with DNA of interest, wherein each of the plurality of cells is located on a separate pallet removably coupled to a plate,
identifying a stable transfectant of a cell from the plurality of cells in less than 20 cell divisions after transfection or microrganism-mediated infection of the plurality of cells, and
collecting the stable transfectant of a cell by dislodging its pallet from the plate.
27. The method of claim 26 wherein the step of identifying includes identifying the stable transfectant of the cell in less than 10 cell divisions after transfection or microrganism-mediated infection of the plurality of cells.
28. The method of claim 26 wherein the cell volume is less than 1 nL.
29. The method of claim 26 wherein the cell volume is less than 100 pL.
30. The method of claim 26 wherein the plurality of cells are arranged in an array.
31. The method of claim 26 wherein the cell is adherent to a surface.