Cell motility assay
View Patent ↗An electrooptical method is disclosed for quantifying the motility response of cells to external stimuli, wherein a target cell population (formed as a function of said response) is subsequently optically differentiated by the addition of an optical differentiation solution, and whereby the targeted cell population becomes more or less detectable to an electrooptical reading device. The method can be used to perform chemotactic assays. For such and other purposes, the method can be performed utilizing high throughput robotic automation.
1. A method for analyzing cells utilizing an electrooptical reading device capable of detecting an optical property of said cells, the method comprising in order the steps of:
(a) providing a porous film having a starting surface and a destination surface;
(b) depositing said cells on the starting surface of said porous film and promoting migration of a portion of said cells from said starting surface to said destination surface through said porous film;
(c) adding an optical differentiation solution to either the portion of the cells remaining on the starting surface or the portion of the cells migrating to the destination surface to render the portion to which said optical differentiation solution was added more or less detectable by said electrooptical reading device; and
(d) reading said optical property of said cells remaining on the starting surface or migrating to the destination surface or both utilizing said electrooptical reading device.
2. The method of claim 1 , wherein the cells placed in step (b) on the starting surface of said porous film are labeled with a compound that emits radiation of a predetermined fluorescent bandwidth upon exposure to radiation of a different predetermined exposure bandwidth.
3. The method of claim 2 , wherein said compound is calcein, a calcein precursor, or a calcein derivative.
4. The method of claim 2 wherein the portion of the cells remaining on the starting surface are rendered less detectable by applying onto the cells that do not migrate to said destination surface and said starting surface said optical differentiation solution, said optical differentiation solution containing a colorant capable of absorbing light at least within said predetermined fluorescent bandwidth, said optical differentiation solution substantially incapable of flowing through said porous film at least for the duration of steps (c) and (d).
5. The method of claim 4 , wherein said optical differentiation solution comprises a suspension of said colorant in glycerol, and wherein said colorant is carbon black.
6. The method of claim 5 , wherein the electrooptical reading device in step (d) is utilized to detect irradiation within said predetermined fluorescent bandwidth upon exposure of said migrant portion of said cells with irradiation of said predetermined exposure bandwidth, said exposure not passing through said porous film prior to incidence upon said migrant portion of cells.
7. The method of claim 1 , wherein the porous film is an isoporous polycarbonate membrane.
8. The method of claim 1 , wherein said porous film is provided between nested top and bottom fluid receptacles, said nested fluid receptacle being divided into an upper zone and a lower zone by said porous film.
9. The method of claim 8 , wherein said method is conducted utilizing a multi-well array, said multi-well array containing an array of said nested fluid receptacles.
10. The method of claim 9 , wherein said multi-well array comprises twenty-four of said nested fluid receptacles.
11. The method of claim 8 , wherein the migration in step (b) of said portion of said cells from said starting surface to said destination surface through said porous film is promoted by placing a chemoattractant-containing solution in said lower zone.
12. The method of claim 11 , wherein the portion of the cells remaining on the starting surface are rendered less detectable by said electrooptical reading device by adding said optical differentiation solution into said upper zone, said optical differentiation solution containing a colorant capable of absorbing light at least within said predetermined fluorescent bandwidth, said solution substantially incapable of flowing through said porous film at least for the duration of steps (c) through (d).