# US 7,408,176 B2 · System and method employing photokinetic techniques in cell biology imaging applications

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- Number: US 7,408,176 B2
- Document ID: US7408176B2
- Status: Granted
- Kind: B2
- Application number: 10/872,329
- Filed: 2004-06-18
- Published: 2005-01-13
- Granted: 2008-08-05
- Earliest claimed: 2003-06-19
- Assignees: Applied Precision, LLC
- Inventors: Paul C. Goodwin; Carl S. Brown; Steven A. Reese
- Web page: https://patentgrep.com/patent/US7408176B2

## CPC

- G01N 33/582: {with fluorescent label}
- G01N 15/1433: using image recognition
- G01N 21/6428: {Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53)}
- G01N 21/6452: {Individual samples arranged in a regular 2D-array, e.g. multiwell plates}
- G01N 21/6458: {Fluorescence microscopy (fluorescence microscopes per se G02B21/0076 and G02B21/16)}
- G01N 33/5005: {involving human or animal cells (immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025)}
- G06T 5/73: Deblurring; Sharpening
- G06T 2207/10064: Fluorescence image
- G06T 2207/30024: Cell structures in vitro; Tissue sections in vitro
- 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)
- G06T: IMAGE DATA PROCESSING OR GENERATION, IN GENERAL

## Abstract

A system and method employing photokinetic techniques in cell biology imaging applications are disclosed. Systems and methods of acquiring image data of an object may comprise: selectively inducing photoactivation of material at a site on the object; performing an optical axis integration scan; simultaneously executing a time delay integration scan sequence; and processing acquired image data in accordance with one or more desired analyses. Various methodologies and applications may include, inter alia, selective photobleaching of a site on the object, diffusion rate, velocity, and wave-front propagation analyses, multi-dimensional analyses of dispersion characteristics, biomolecular binding in cellular organelles, and photoactivation assisted systematic image segmentation for the study of cellular components.

## Claims

### Claim 1

A method of acquiring image data of an object; said method comprising:
selectively inducing photoactivation of material at a site on the object;
performing an optical axis integration scan;
simultaneously executing a time delay integration scan sequence; and
selectively repeating said performing and said executing.

### Claim 2 (depends on 1)

The method of claim 1 wherein said performing comprises acquiring image data of the object at an image plane positioned along an optical axis.

### Claim 3 (depends on 2)

The method of claim 2 wherein said performing further comprises providing relative translation along said optical axis of the object and said image plane.

### Claim 4 (depends on 3)

The method of claim 3 wherein said executing comprises providing relative translation along a lateral axis of the object and said image plane.

### Claim 5 (depends on 4)

The method of claim 4 wherein said executing comprises synchronizing said relative translation along said lateral axis with a data acquisition rate associated with an imaging device.

### Claim 6 (depends on 3)

The method of claim 3 wherein said performing further comprises selectively alternating a direction of said relative translation along said optical axis.

### Claim 7 (depends on 2)

The method of claim 2 wherein said performing further comprises integrating said image data concomitantly with said acquiring.

### Claim 8 (depends on 7)

The method of claim 7 further comprising deblurring said image data subsequent to said integrating.

### Claim 9 (depends on 7)

The method of claim 7 further comprising deconvolving said image data subsequent to said integrating.

### Claim 10 (depends on 1)

The method of claim 1 wherein said selectively inducing comprises tagging the object to be scanned with a Green Fluorescent Protein variant.