# US 10,369,378 B2 · System for optical stimulation of target cells

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 10,369,378 B2
- Document ID: US10369378B2
- Status: Granted
- Kind: B2
- Application number: 16/126,895
- Filed: 2018-09-10
- Published: 2019-04-11
- Granted: 2019-08-06
- Earliest claimed: 2007-01-10
- Assignees: The Board of Trustees of the Leland Stanford Junior University
- Applicants: The Board of Trustees of the Leland Stanford Junior University
- Inventors: Karl Deisseroth; Feng Zhang; Edward Boyden
- Web page: https://patentgrep.com/patent/US10369378B2

## CPC

- A61N 5/06: using light
- A61N 5/0601: {Apparatus for use inside the body}
- A61N 5/0613: {Apparatus adapted for a specific treatment}
- A61N 5/0622: {Optical stimulation for exciting neural tissue}
- C12N 5/0602: {Vertebrate cells}
- C12N 13/00: Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
- G01N 33/502: {for testing non-proliferative effects}
- G01N 33/6872: {Intracellular protein regulatory factors and their receptors, e.g. including ion channels}
- A61N 2005/0626: {Monitoring, verifying, controlling systems and methods}
- A61N 2005/0647: {the applicator adapted to be worn on the head}
- A61N 2005/0652: {Arrays of diodes}
- A61N 2005/0663: {Coloured light}
- A61N 2005/0665: {Reflectors}
- A61N: ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY (measurement of bioelectric currents A61B; surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body A61B18/00; anaesthetic apparatus in general A61M; incandescent lamps H01K; infrared radiators for heating H05B)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

Various systems and methods are implemented for controlling stimulus of a cell. One such method is implemented for optical stimulation of a cell expressing an NpHR ion pump. The method includes the step of providing a sequence of stimuli to the cell. Each stimulus increases the probability of depolarization events occurring in the cell. Light is provided to the cell to activate the expressed NpHR ion pump, thereby decreasing the probability of depolarization events occurring in the cell.

## Claims

### Claim 1

A method for screening a candidate agent that modulates ion channel function in a mammalian cell, the method comprising:
(i) expressing in a mammalian cell a light-driven chloride ion pump from Natronobacterium pharaonis (NpHR);
(ii) expressing in the mammalian cell an ion channel;
(iii) labeling the mammalian cell with a voltage sensitive dye;
(iv) exposing the mammalian cell to a first light;
(v) recording a first fluorescence signal of the voltage sensitive dye;
(vi) exposing the mammalian cell expressing NpHR to the candidate agent;
(vii) exposing the mammalian cell expressing NpHR to a second light; and
(viii) recording the second fluorescence signal of the voltage sensitive dye, and
wherein if the second fluorescence signal of the voltage sensitive dye is greater than the first fluorescence signal of the voltage sensitive dye, then the candidate agent facilitates ion channel function.

### Claim 2 (depends on 1)

The method of claim 1, wherein if the first fluorescence signal of the voltage sensitive dye is equal to the second fluorescence signal of the voltage sensitive dye, the candidate agent does not have an affect ion channel function.

### Claim 3 (depends on 1)

The method of claim 1, wherein if the second fluorescence signal of the voltage sensitive dye is smaller than the first fluorescence signal of the voltage sensitive dye, than the candidate agent diminishes ion channel function.

### Claim 4 (depends on 1)

The method of claim 1, wherein the ion channel is a Channelrhodopsin-2 (ChR2) ion channel.

### Claim 5 (depends on 4)

The method of claim 4, wherein the first light is a yellow light or a blue light, wherein the yellow light activates the NpHR ion pump expressed in the mammalian cell and the blue light activates the ChR2 ion channel expressed in the mammalian cell.

### Claim 6 (depends on 1)

The method of claim 1, wherein the voltage sensitive dye is a calcium-sensitive dye.

### Claim 7 (depends on 1)

The method of claim 1, wherein the second light is a yellow light or a blue light, wherein the yellow light activates the NpHR ion pump expressed in the mammalian cell and the blue light activates the ChR2 ion channel expressed in the mammalian cell.