Use of live cell inteferometry with reflective floor of observation chamber to determine changes in mass of mammalian cells

• US 9,810,683 B2
• Filed: 11/25/2013
• Issued: 11/07/2017
• Est. Priority Date: 05/06/2009
• 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: Gimzewski, James K., Reed, Jason C., Teitell, Michael A.
• Primary Examiner(s): LEE, HWA S
• Application Number:
  US14/088,992
• Publication Number:
  US 20140080171A1
• Time in Patent Office: 1,443 Days
• CPC Class Codes: G01J 3/453 by correlation of the amplitudes
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G02B 21/0056 based on optical coherence, e.g. phase-contrast arrangements, interference arrangements
• CPC Class Codes: G02B 21/32 Micromanipulators structurally combined with microscopes
• CPC Class Codes: G02B 5/126 including curved refracting surface

Abstract

The present invention provides optical systems and methods for determining a characteristic of a cell, such as cell type, cellular response to a biochemical event, biological state and the like. The methods typically involve using interferometry to observe membrane properties in a cell and then use this information to determine one or more characteristics of a cell. The methods of the invention are useful for applications such as drug screening as well as diagnostic techniques.

Claims

1. A method for determining changes in optical thickness and/or mass of a mammalian cell, the method comprising:

(a) placing a mammalian cell in an observation chamber of a microscope, wherein the observation chamber is adapted to form an aqueous environment, and wherein said observation chamber comprises a reflective floor;

(b) using an interference microscope to detect changes in optical path length introduced by said cell as light passes through said cell and is reflected back from said reflective floor;

(c) altering the first environment to form a second environment;

(d) using said interference microscope to detect changes in optical path length introduced by said cell as light passes through said cell in said second environment and is reflected back from said reflective floor;

(e) comparing the observations in (b) with the observations in (d) so as to determine changes in the optical thickness and/or mass of said cell resulting from the alteration of said first environment to said second environment.

2. The method of claim 1, wherein the method comprises determining changes in the optical thickness of said mammalian cell.

3. The method of claim 1, wherein said method comprises determining changes in mass of said mammalian cell that occur following combining said mammalian cell with a composition comprising a biologically active agent.

4. The method of claim 3, wherein the method is used to determine a response to stimuli in a plurality of mammalian cells.

5. The method of claim 1, wherein the method is used to determine changes in the mass of said mammalian cell under changing environmental conditions over time.

6. The method of claim 1, wherein said microscope detects changes in optical path length as small as 1 nanometer.

7. The method of claim 1, wherein the method is performed using a system comprising:

(a) a detector operatively coupled to said interference microscope;

(b) a sample assembly comprising the observation chamber, wherein said observation chamber has a reflective floor;

(c) a reference assembly comprising a reference chamber adapted to contain a fluid;

(d) a memory storage element adapted to store one or more images of the cell; and

(e) a processor element adapted to process one or more images of the cell.

8. The method of claim 7, wherein said interference microscope includes a reference assembly comprising:

a first optical window;

a first housing element adapted to hold the first optical window;

a second optical window;

a second housing element adapted to hold the second optical window; and

a plurality of spacer elements disposable between the first optical window and the second optical window and adapted to separate the first and second optical windows to a defined distance.

9. The method of claim 7, wherein the method comprises obtaining a plurality of images of said cell and storing said images in the memory storage element.

10. The method of claim 1, further comprising removing the cell from the observation chamber and manipulating the cell for further analysis.

11. The method of claim 1, wherein said interference microscope comprises a Michelson interferometer.