Single cell drug response measurements via live cell interferometry

• US 10,203,331 B2
• Filed: 08/02/2012
• Issued: 02/12/2019
• Est. Priority Date: 08/02/2011
• Status: Active Grant

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Reed, Jason C., Teitell, Michael A.
• Primary Examiner(s): Vanni, G Steven
• Application Number:
  US14/235,547
• Publication Number:
  US 20140178865A1
• Time in Patent Office: 2,385 Days
• Field of Search: None
• CPC Class Codes: G01B 11/0675 using interferometry
• CPC Class Codes: G01B 9/02057 by using common path configuration, i.e. reference and object path almost entirely overlapping
• CPC Class Codes: G01B 9/02089 Displaying the signal, e.g. for user interaction
• CPC Class Codes: G01B 9/0209 Low-coherence interferometers
• CPC Class Codes: G01N 33/502 for testing non-proliferative effects
• CPC Class Codes: G01N 33/5026 on cell morphology
• CPC Class Codes: G01N 33/57492 involving compounds localized on the membrane of tumor or cancer cells

Abstract

A central question in cancer therapy is how individual cells within a population of tumor cells respond to drugs designed to arrest their growth. However, the absolute growth of cells, their change in physical mass, whether cancerous or physiologic, is difficult to measure directly with traditional techniques. Embodiments of the invention provide live cell interferometry (LCI) for rapid, realtime quantification of cell mass in cells exposed to a changing environment. Overall, LCI provides a conceptual advance for assessing cell populations to identify, monitor, and measure single cell responses, such as to therapeutic drugs.

Claims

1. A method for measuring a cellular response to an environment comprising:

(a) placing a test cell in a first environment;

(b) using an interference microscope to perform live cell interferometry and thereby determine a measure proportional to the mass of said test cell, where said live cell interferometry comprises measuring the fractional phase shift between a test beam of light propagating through said test cell and a reference beam of light with said interference microscope and integrating said phase shift over the area of said test cell to provide said measure proportional to the mass of said test cell;

(c) placing a reference cell in a second environment;

(d) using an interference microscope to perform live cell interferometry and thereby determine a measure proportional to the mass of said reference cell, where said live cell interferometry comprises measuring the fractional phase shift between a test beam of light propagating through said reference cell and a reference beam of light with said interference microscope and integrating said phase shift over the area of said reference cell to provide said measure proportional to the mass of said reference cell; and

(e) determining the difference between the measure determined in (b) and the measure determined in (d) to thereby provide a measure of the change in mass of said test cell in response to the first environment.

2. The method of claim 1, wherein said test cell is present in the first environment as an isolated single cell.

3. The method of claim 1, wherein said test cell is present in the first environment in a cluster or clump of cells.

4. The method of claim 1, wherein mass properties of a plurality of test cells present in the first environment are observed.

5. The method of claim 1, wherein the first environment comprises a test composition and the second environment does not comprise the test composition.

6. The method of claim 5, wherein the test composition comprises an antibiotic, an antibody, an alkylating agent, an antimetabolite, a cell cycle inhibitor, a topoisomerase inhibitor, an siRNA or a cell.

7. The method of claim 6, wherein the test composition comprises an antibody that binds HER2.

8. The method of claim 5, wherein said method comprises determining the measure proportional to the mass of the test cell in the first environment a plurality of times to thereby measure changes in the mass property of the cell over a period of time.

9. The method of claim 8, wherein changes in said measure proportional to the mass of the test cell are observed over time to thereby determine a temporal mass profile.

10. The method of claim 9, further comprising comparing said temporal mass profile to a database of temporal mass profiles, wherein the database of temporal mass profiles is selected to include temporal mass profiles that are characteristic of cellular sensitivity to the test composition and temporal mass profiles that are characteristic of cellular resistance to the test composition.

11. The method of claim 1, wherein the mass of the test cell is determined from said measure proportional to the mass of a test cell using an equation:

m=i/α

∫

φ

λ

dA

wherein m is the mass of the cell, a is a constant describing a relationship between the phase shift and cell mass, φ

is the measured fractional phase shift, λ

is the illumination wavelength, and integration is performed across an entire cell area, A.

12. The method of claim 11, wherein α

=1.8×

10<sup>−

3</sup> m³ kg<sup>−

1</sup>.

13. The method of claim 11, wherein said interference microscope comprises:

a detector operatively coupled to the microscope;

a sample assembly comprising an observation chamber that contains said test cell;

a reference assembly comprising a reference chamber that contains said reference cell; and

a beam splitter for splitting a light beam from a light source into a test beam and a reference beam.

14. The method of claim 13, wherein the observation chamber comprises at least one perfusion conduit that circulates a cell media within the observation chamber.