High Resolution Label Free Analysis of Cellular Properties

• US 20090305304A1
• Filed: 04/09/2009
• Published: 12/10/2009
• Est. Priority Date: 04/09/2008
• Status: Active Grant

• Current Assignee: X-Body Incorporated (Bristol-Myers Squibb Company)
• Original Assignee: Sru Biosystems Incorporated
• Inventors: Laing, Lance G., Wagner, Rick, Fernandez, Rafael
• Granted Patent: US 8,257,936 B2
• Time in Patent Office: Days
• US Class Current: 435/7.200
• CPC Class Codes: G01N 21/4788 Diffraction for sizing particles G01N15/0205
• CPC Class Codes: G01N 21/7703 using reagent-clad optical fibres or optical waveguides using measurement of total internal reflection or attenuated total reflection G01N21/552; optical fibres or waveguides per se G02B
• CPC Class Codes: G01N 33/5026 on cell morphology
• CPC Class Codes: G01N 33/5029 on cell motility
• CPC Class Codes: G01N 33/5032 on intercellular interactions
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings

Abstract

The invention provides methods of detecting a change in cell growth patterns.

Claims

1. A method of detecting a change in a cell growth pattern comprising:

a) immobilizing one or more cells to a location on a surface of a calorimetric resonant reflectance optical biosensor with an antibody or a binding fragment thereof that specifically binds an adhesion protein;

b) detecting a calorimetric resonant reflectance optical peak wavelength value (PWV) for the location;

c) incubating the one or more cells for a period of time or applying a test reagent to the one or more cells and incubating the one or more cells for a period of time;

d) detecting the calorimetric resonant reflectance optical PWV for the location; and

e) comparing the PWV of step (b) to the PWV of step (d);

wherein a difference between the colorimetric resonant reflectance optical PWV of step (b) in relation to the colorimetric resonant reflectance optical PWV of step (d) indicates a change in the cell growth pattern in the one or more cells.

2. The method of claim 1 wherein the change in cell growth pattern is a change in cell morphology, change in cell adhesion, change in cell migration, change in cell proliferation, change in cell death, change in microtubule structure, change in microfilament structure, granule exocytosis, respiratory burst, cell differentiation, or a combination thereof.

3. The method of claim 1, wherein the PWV is detected using a scanner with a lens having a lower limit pixel size of about 1 micrometers to about 15 micrometers.

4. The method of claim 1, wherein the location on a surface of a colorimetric resonant reflectance optical biosensor is an internal surface of a vessel selected from the group consisting of a microtiter well, microtiter plate, test tube, Petri dish, microfluidic channel, and microarray.

5. The method of claim 1, wherein the method is completed in less than one hour.

6. A method of detecting a change in a cell growth pattern comprising:

a) adding one or more cells to a colorimetric resonant reflectance optical biosensor, wherein one or more antibodies or a binding fragments thereof that specifically bind one or more adhesion proteins are immobilized on a location on the biosensor surface;

b) optionally adding one or more adhesion proteins that specifically bind the one or more antibodies or binding fragments thereof to the biosensor surface;

c) detecting a first calorimetric resonant reflectance optical peak wavelength value (PWV) for the location;

d) incubating the one or more cells for a period of time or applying a test reagent to the one or more cells and incubating the one or more cells for a period of time;

e) detecting a second PWV for the location; and

f) comparing the first PWV and second PWV;

wherein a difference between the first PWV in relation to the second PWV indicates a change in the cell growth pattern in the one or more cells.

7. The method of claim 6, wherein the change in cell growth pattern is a change in cell morphology, change in cell adhesion, change in cell migration, change in cell proliferation, change in cell death, change in microtubule structure, change in microfilament structure, granule exocytosis, respiratory burst, cell differentiation, or a combination thereof.

8. The method of claim 6, wherein the first and second PWVs are detected using a scanner with a lens having a lower limit pixel size of about 1 micrometers to about 15 micrometers.

9. The method of claim 6, wherein the location on a surface of a calorimetric resonant reflectance optical biosensor is an internal surface of a vessel selected from the group consisting of a microtiter well, microtiter plate, test tube, Petri dish, microfluidic channel, and microarray.

10. The method of claim 6, wherein the method is completed in less than one hour.

11. The method of claim 6, wherein the one or more cells expresses an adhesion protein that specifically binds the one or more antibodies or binding fragments thereof.

12. The method of claim 6, wherein the one or more cells expresses a receptor that specifically binds the one or more adhesion proteins.

13. A method of detecting a change in a cell growth pattern comprising:

a) adding one or more cells to a calorimetric resonant reflectance optical biosensor, wherein one or more antibodies or a binding fragments thereof that specifically bind one or more adhesion proteins are immobilized on a location on the biosensor surface;

b) detecting calorimetric resonant reflectance optical peak wavelength values (PWVs) for the location over the whole time of the assay;

b) optionally adding one or more adhesion proteins that specifically bind the one or more antibodies or binding fragments thereof to the biosensor surface;

d) incubating the one or more cells for a period of time or applying a test reagent to the one or more cells and incubating the one or more cells for a period of time;

wherein a difference between the PWVs over the time of the assay indicates a change in the cell growth pattern in the one or more cells.

14. The method of claim 13, wherein the change in cell growth pattern is a change in cell morphology, change in cell adhesion, change in cell migration, change in cell proliferation, change in cell death, change in microtubule structure, change in microfilament structure, granule exocytosis, respiratory burst, cell differentiation, or a combination thereof.

15. The method of claim 13, wherein the first and second PWVs are detected using a scanner with a lens having a lower limit pixel size of about 1 micrometers to about 15 micrometers.

16. The method of claim 13, wherein the location on a surface of a calorimetric resonant reflectance optical biosensor is an internal surface of a vessel selected from the group consisting of a microtiter well, microtiter plate, test tube, Petri dish, microfluidic channel, and microarray.

17. The method of claim 13, wherein the method is completed in less than one hour.

18. The method of claim 13, wherein the one or more cells expresses an adhesion protein that specifically binds the one or more antibodies or binding fragments thereof.

19. The method of claim 13, wherein the one or more cells expresses a receptor that specifically binds the one or more adhesion proteins.