Optical cytometry

• US 8,599,383 B2
• Filed: 05/06/2009
• Issued: 12/03/2013
• 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: Teitell, Michael A., Gimzewski, James K., Reed, Jason C.
• Primary Examiner(s): LEE, HWA S
• Application Number:
  US12/436,702
• Publication Number:
  US 20100284016A1
• Time in Patent Office: 1,672 Days
• Field of Search: 356/517, 356/450, 356/481, 356/496, 356/337, 356/338, 356/341, 356/342, 356/343
• US Class Current: 356/451
• 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 system for obtaining an image of a cell comprising:

(a) microscope capable of measuring a feature of interest in a sample;

(b) a detector operatively coupled to the microscope;

(c) a sample assembly comprising an observation chamber adapted to contain the cell, wherein the observation chamber comprises at least one perfusion conduit adapted to circulate a cell media within the chamber;

(d) 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; and

(e) a plurality of reflective microparticles capable of adhering to the cell, wherein the average diameter of the reflective microparticles is between 0.5 μ

m and 30 μ

m.

2. The system of claim 1, wherein the microscope is a confocal microscope.

3. The system of claim 1, wherein the sample assembly further comprises:

a viewing window;

a first housing element adapted to hold the viewing window;

and wherein the thickness of the viewing window is equivalent to the combined thickness of the first and second optical windows in the reference assembly.

4. The system of claim 1, wherein the reflective microparticles are spherical magnetic microparticles having an average diameter of between 1 μ

m and 15 μ

m.

5. The system of claim 1, wherein the reflective microparticles comprise a gradient index (GRIN) spherical lens.

6. The system of claim 1, further comprising a magnet disposed below the observation chamber and oriented coaxially with an optical axis.

7. The system of claim 6, wherein the magnet is operatively coupled to a motorized micrometer and adapted to exert a magnetic force of between 0 Newtons and 5 nanoNewtons on a magnetic reflective microparticle adhered to the surface of the cell.

8. The system of claim 6, wherein the magnet is adapted to generate a magnetic field of between 200 Gauss and 3 kiloGauss and/or a magnetic field gradient range of between 300,000 to 800,000 Gauss/meter.

9. The system of claim 6, wherein magnet is operatively coupled to the system so as to allow the measurement of an axial position of a magnetic reflective microparticle in a sample.

10. The system of claim 6, wherein magnet is operatively coupled to a motorized micrometer configured to allow the measurement of a z motion of a magnetic reflective microparticle in a sample.

11. The system of claim 1, further comprising a processor element and a memory storage element adapted to process and store one or more images of the cell.

12. A system for obtaining an image of a cell comprising:

(a) a interference microscope capable of extracting information from interferometric fringes;

(b) a detector operatively coupled to the interference microscope;

(c) a sample assembly comprising an observation chamber adapted to contain the cell, wherein the observation chamber comprises at least one perfusion conduit adapted to circulate a cell media within the chamber;

(d) a reference assembly adapted to substantially match an optical path length of the sample assembly,

(e) a plurality of reflective microparticles capable of adhering to the cell, wherein the average diameter of the reflective microparticles is between 0.5 μ

m and 30 μ

m; and

a magnet disposed below the observation chamber and oriented coaxially with an optical axis, wherein the magnet is operatively coupled to a motorized micrometer configured to allow the measurement of a z motion of the microparticle in a sample.

13. A method for observing a property of a cell, the method comprising:

(a) adhering a reflective magnetic microparticle to the cell;

(b) placing the cell in a observation chamber of a interference microscope system capable of extracting information from interferometric fringes, wherein;

the observation chamber comprises at least one perfusion conduit adapted to circulate a cell media within the chamber; and

a magnet is disposed below the observation chamber and oriented coaxially with an optical axis, wherein the magnet is operatively coupled to a motorized micrometer configured to allow the measurement of a z motion of the microparticle in a sample;

(c) exposing the cell coated with the microparticle to a magnetic field using the magnet disposed below the observation chamber; and

(d) using the microscope to observe the movement of the microparticle adhered to the cell in response to the applied magnetic field, wherein the movement of the reflective microparticle adhered to the cell correlates to a property of the cell, so that a property of the cell is observed.

14. A system for obtaining an image of a cell comprising:

(a) a microscope capable of measuring a feature of interest in a sample;

(b) a detector operatively coupled to the microscope;

(c) a sample assembly comprising an observation chamber adapted to contain the cells wherein the observation chamber comprises at least one perfusion conduit adapted to circulate a cell media within the chamber;

(d) a plurality of magnetic reflective microparticles capable of adhering to the cell, wherein the average diameter of the magnetic microparticles is between 0.5 btm and 30 btm; and

(e) a magnet disposed below the observation chamber and oriented coaxially with an optical axis, wherein the magnet is operatively coupled to a motorized micrometer and adapted to exert a magnetic force of between 0 Newtons and 5 nanoNewtons on magnetic reflective microparticles adhered to the surface of the cell.