# US 9,040,272 B2 · Apparatus and method for living cell manipulation

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- Number: US 9,040,272 B2
- Document ID: US9040272B2
- Status: Granted
- Kind: B2
- Application number: 13/158,843
- Filed: 2011-06-13
- Published: 2011-12-15
- Granted: 2015-05-26
- Earliest claimed: 2010-06-14
- Assignees: Hamilton Thorne, Inc.
- Applicants: Diarmaid H. Douglas-Hamilton; Thomas G. Kenny
- Inventors: Diarmaid H. Douglas-Hamilton; Thomas G. Kenny
- Web page: https://patentgrep.com/patent/US9040272B2

## CPC

- C12N 13/00: Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
- C12M 35/02: {Electrical or electromagnetic means, e.g. for electroporation or for cell fusion}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS}

## Abstract

The present invention provides a method and system for using eye-safe infrared energy from a Class I laser to manipulate cells in culture. The laser energy produces one or more phase boundary propulsion events, which generate hydrodynamic forces sufficient to manipulate cells at the focal point.

## Claims

### Claim 1

A method of repositioning one or more of a plurality of cells suspended in an aqueous medium by inducing phase boundary propulsion (PBP) at a focal point in the aqueous medium, comprising providing infrared energy having a wavelength greater than 1400 nm and an energy less than 1.6 mJ per pulse from a laser through an objective lens to the aqueous medium, superheating the aqueous medium and inducing a phase transition at a temperature insufficient to produce significant ionization, moving the focal point through the medium, and repeating said step of providing the infrared energy, to produce a series of adjacent PBP bubbles.

### Claim 2 (depends on 1)

The method of claim 1, wherein said step of providing comprises providing repeated pulses at 100 to 1000 Hz.

### Claim 3 (depends on 2)

The method of claim 2, wherein said step of providing comprises providing the repeated pulses at about 100 to 200 Hz.

### Claim 4 (depends on 1)

The method of claim 1, wherein said step of providing comprises providing the infrared energy in at least two sequential pulses having different wavelengths, producing separate superheating and phase transition events.

### Claim 5 (depends on 1)

The method of claim 1, wherein said step of providing comprises providing the infrared energy in one or more pulses of duration less than 4 ms.

### Claim 6 (depends on 1)

The method of claim 1, wherein said step of providing comprises providing the infrared energy in one or more pulses of duration between 50 and 500 μs.

### Claim 7 (depends on 1)

The method of claim 1, wherein said step of providing comprises providing the infrared energy in one or more pulses of duration of about 150 μs.

### Claim 8 (depends on 1)

The method of claim 1, wherein said step of providing comprises providing the laser energy in repeated pulses having a mean power of less than 90 mW.

### Claim 9 (depends on 4)

The method of claim 4, wherein said sequential pulses are provided from separate lasers.

### Claim 10 (depends on 4)

The method of claim 4, wherein said sequential pulses comprise a first pulse having a wavelength between 1450 and 1490 nm and a second pulse having a wavelength between 1.9 and 2.1 μm, or between 1410 and 1430 nm.

### Claim 11 (depends on 10)

The method of claim 10, wherein the first pulse has a wavelength of about 1450 nm.

### Claim 12 (depends on 10)

The method of claim 10, wherein the second pulse has a wavelength of about 1.9 μm.

### Claim 13 (depends on 10)

The method of claim 10, wherein the second pulse has a wavelength of about 1420 nm.

### Claim 14 (depends on 1)

The method of claim 1, wherein said moving the focal point comprises moving a dish containing the plurality of cells suspended in the aqueous medium on a computerized microscope stage programmed to follow a specified pattern.