Systems, methods, and workflows for optogenetics analysis

• US 10,161,937 B2
• Filed: 07/10/2017
• Issued: 12/25/2018
• Est. Priority Date: 08/23/2010
• Status: Active Grant

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Inventors: Cohen, Adam Ezra, Kralj, Joel, Douglass, Adam D., Hochbaum, Daniel
• Primary Examiner(s): Navarro, Albert M
• Application Number:
  US15/645,426
• Publication Number:
  US 20180031553A1
• Time in Patent Office: 533 Days
• CPC Class Codes: A61K 49/0017 Fluorescence in vivo
• CPC Class Codes: A61K 49/0097 Cells, viruses, ghosts, red blood cells, viral vectors, used for imaging or diagnosis in vivo
• CPC Class Codes: G01N 2333/705 Assays involving receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: G01N 33/566 using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds

Abstract

The invention provides methods for characterizing cellular physiology by incorporating into an electrically excitable cell an optical reporter of, and an optical actuator of, electrical activity. A signal is obtained from the optical reporter in response to a stimulation of the cell. Either or both of the optical reporter and actuator may be based on genetically-encoded rhodopsins incorporated into the cell. The invention provides all optical methods that may be used instead of, or as a complement to, traditional patch clamp technologies and that can provide rapid, accurate, and flexible assays of cellular physiology.

Claims

1. A method for measuring network effects, the method comprising:

using an optical actuator of electrical activity incorporated into a first plurality of cells of one cell type;

obtaining a signal using an optical reporter of electrical activity incorporated into a second plurality of cells of another cell type; and

evaluating the signal.

2. The method of claim 1, wherein the first plurality of cells of one cell type further comprises an optical reporter of electrical activity.

3. The method of claim 2, wherein the second plurality of cells of one cell type further comprises an optical actuator of electrical activity.

4. The method of claim 1, wherein the first plurality of cells comprises a first class of neurons.

5. The method of claim 1, wherein the second plurality of cells comprises a second class of neurons.

6. The method of claim 1, wherein the first plurality of cells comprises a first class of cardiomyocytes.

7. The method of claim 1, wherein the second plurality of cells comprises a second class of cardiomyocytes.

8. The method of claim 1, wherein the first plurality of cells comprises a first class of cells of iPSC'"'"'s.