Image based quantitation of molecular translocation

• US 8,150,136 B2
• Filed: 03/16/2005
• Issued: 04/03/2012
• Est. Priority Date: 03/16/2004
• Status: Active Grant

• Current Assignee: Cytek Biosciences Inc
• Original Assignee: Amnis Corporation (DiaSorin S.p.A.)
• Inventors: George, Thaddeus C., Basiji, David A., Frost, Keith, Hall, Brian E., Ortyn, William E., Seo, Michael J.
• Primary Examiner(s): Velez, Roberto
• Application Number:
  US10/593,018
• Publication Number:
  US 20090202130A1
• Time in Patent Office: 2,575 Days
• Field of Search: None
• US Class Current: 382/133
• CPC Class Codes: G01N 33/56966 Animal cells

Abstract

The use of an imaging system, cell compartment markers, and molecular markers in methods for correlating the movement of molecules within a cell to a particular compartment are provided, including measuring and correlating molecule movement in adherent and non-adherent cells.

Claims

1. A method for measuring molecular movement in a cell, comprising:

contacting a cell with a compartment marker and with a molecular marker;

imaging the marked cell with a detector;

creating a compartment mask; and

correlating said compartment mask and said molecular marker to measure molecular movement in a cell.

2. The method of claim 1 wherein there is relative motion between the cell and the detector.

3. The method of claim 1 wherein the molecular marker is a fluorescent labeled antibody.

4. The method of claim 1 wherein the compartment marker is a fluorescent molecule.

5. The method of claim 1 wherein the compartment marker is a nucleus, cytoplasm, or a membrane.

6. The method of claim 1 wherein the molecule marked is NF-κ

B.

7. The method of claim 1 further comprising the step of inducing molecular movement in the cell.

8. The method of claim 7 wherein the induced molecular movement is nuclear translocation.

9. The method of claim 7 wherein the molecular movement is induced with LPS or IL-1β

/TNF-α

.

17. The method according to any one of claims 1-16 wherein the images are collected simultaneously.

18. The method according to any one of claims 1-16 wherein the detector is a time delay integration charge-coupled detector.

10. A method for measuring nuclear translocation in a cell, comprising:

contacting a cell with a nuclear marker and with a molecular marker;

imaging the marked cell with a detector, creating a nuclear mask; and

correlating said nuclear mask and said molecular marker to measure molecular movement in a cell.

11. The method of claim 10 wherein there is relative motion between the cell and the detector.

12. The method of claim 10 further comprising the step of inducing molecular movement in the cell.

13. The method of claim 12 wherein the induced molecular movement is nuclear translocation.

14. The method of claim 12 wherein the molecular movement is induced with LPS or IL-1β

/TNF-α

.

15. The method of claim 10 wherein the nuclear marker is 7-AAD.

16. The method of claim 10 wherein the molecule marked is NF-κ

B.