High resolution label free analysis of cellular properties

• US 8,257,936 B2
• Filed: 04/09/2009
• Issued: 09/04/2012
• Est. Priority Date: 04/09/2008
• Status: Active Grant

• Current Assignee: X-Body Incorporated (Bristol-Myers Squibb Company)
• Original Assignee: X-Body Incorporated (Bristol-Myers Squibb Company)
• Inventors: Laing, Lance G., Fernandez, Rafael, Wagner, Rick
• Primary Examiner(s): Chin, Chris L
• Application Number:
  US12/421,294
• Publication Number:
  US 20090305304A1
• Time in Patent Office: 1,244 Days
• Field of Search: None
• US Class Current: 435/7.2
• 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 21/7743 the reagent-coated grating coupling light in or out of the waveguide
• 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) adding one or more cells that express one or more adhesion proteins on their surface to a colorimetric resonant reflectance optical biosensor, wherein one or more antibodies or binding fragments thereof are immobilized on one or more locations on the biosensor surface, wherein the antibodies or binding fragments thereof specifically bind to the one or more adhesion proteins;

b) detecting a first colorimetric resonant reflectance optical peak wavelength value (PWV) for the one or more locations;

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 a second PWV for the one or more locations; and

e) 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.

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 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.

4. The method of claim 1, wherein the one or more locations on a surface of a colorimetric resonant reflectance optical biosensor are present on 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, 45 minutes, 30 minutes, 20 minutes, 10 minutes, 5 minutes, or 3 minutes.

6. The method of claim 1, wherein the method is completed in less than 8 hours, 7 hours, 6 hours, 5 hours, 4 hours, 3 hours or 2 hours.

7. The method of claim 1, wherein the cells are monitored in real time over days.

8. The method of claim 1, wherein colorimetric resonant reflectance optical peak wavelength values (PWVs) are determined for one or more locations over the whole time of the assay 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.