Method for acquisition, storage, and retrieval of cell screening data on a computer system

• US 7,565,247 B1
• Filed: 11/22/2000
• Issued: 07/21/2009
• Est. Priority Date: 02/27/1997
• Status: Expired due to Term

• Current Assignee: Cellomics Incorporated (Thermo Fisher Scientific Incorporated)
• Original Assignee: Cellomics Incorporated (Thermo Fisher Scientific Incorporated)
• Inventors: Dunlay, R. Terry, Taylor, D. Lansing
• Primary Examiner(s): Smith; Carolyn L.
• Application Number:
  US09/718,770
• Time in Patent Office: 3,163 Days
• Field of Search: 702/19, 435/4, 435/6, 422/50, 703/11
• US Class Current: 702/19
• CPC Class Codes: B01J 2219/00315 Microtiter plates
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00707 separated from the reactor apparatus
• CPC Class Codes: B01J 2219/00743 Cells
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/6813 Hybridisation assays
• CPC Class Codes: C12Q 2525/179 incorporating arbitrary or random nucleotide sequences
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 15/1433 using image recognition
• CPC Class Codes: G01N 2015/1486 Counting the particles
• CPC Class Codes: G01N 2015/1497 Particle shape
• CPC Class Codes: G01N 2021/6419 Excitation at two or more wavelengths
• CPC Class Codes: G01N 2021/6482 Sample cells, cuvettes
• CPC Class Codes: 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
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 33/5005 involving human or animal cells immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025
• CPC Class Codes: G01N 33/5035 on sub-cellular localization
• CPC Class Codes: G01N 33/582 : with fluorescent label
• CPC Class Codes: G02B 21/16 : adapted for ultraviolet illumination ; Fluorescence microscopes G02B21/0076 takes precedence
• CPC Class Codes: G02B 21/365 : Control or image processing arrangements for digital or video microscopes G02B21/361, G02B21/362 take precedence
• CPC Class Codes: Y10S 436/80 : Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10S 436/807 : Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/809 : Multifield plates or multicontainer arrays
• CPC Class Codes: Y10S 977/795 : Composed of biological material
• CPC Class Codes: Y10S 977/881 : Microscopy or spectroscopy, e.g. sem, tem
• CPC Class Codes: Y10S 977/92 : Detection of biochemical
• CPC Class Codes: Y10S 977/941 : including dna logic element
• CPC Class Codes: Y10S 977/942 : including protein logic element

Abstract

The invention relates to an optical system for determining the distribution, environment, or activity of fluorescently labeled reporter molecules in cells for the purpose of screening large numbers of compounds for specific biological activity. The invention involves providing cells containing fluorescent reporter molecules in an array of locations and scanning numerous cells in each location with a fluorescent microscope, converting the optical information into digital data, and utilizing the digital data to determine the distribution, environment or activity of the fluorescently labeled reporter molecules in the cells. The array of locations may be an industry standard 96 well or 384 well microtiter plate or a microplate which is a microplate having a cells in a micropaterned array of locations. The invention includes apparatus and computerized method for processing, displaying and storing the data.

Claims

1. A method for acquisition, storage, and retrieval of cell screening data on a computer system, comprising the steps of:

a) providing a plate containing wells, wherein the wells comprise cells;

b) storing input parameters used for screening of the plate in a computer system database;

c) repeating steps (i)-(ix) for a desired number of wells;

i) selecting an individual well on the plate,

ii) collecting subcellular image data from the cells in the well,

iii) storing the subcellular image data in the computer system database,

iv) collecting feature data from the subcellular image data,

v) storing the feature data in the computer system database,

vi) calculating well summary data using the subcellular image data and the feature data collected from the well;

vii) storing the well summary data in the computer system database;

viii) calculating plate summary data using the well summary data from the computer system database; and

ix) storing the plate summary data in the computer system database;

wherein the subcellular image data, the feature data, the well summary data, and the plate summary data can be retrieved from the computer system database.

2. A computer readable medium having stored therein instructions for causing a computer to execute the method of claim 1.

3. The method of claim 1 wherein the wells include cells treated with a test compound.

4. The method of claim 1 wherein the plate comprises a microplate.

5. The method of claim 1 wherein the computer system database includes microplate data.

6. The method of claim 1 wherein the computer system database includes photographic subcellular image data.

7. The method of claim 1 wherein the input parameters used for screening of the plate include parameters for one or more of the following:

identifying nuclei;

identifying cytoplasm;

identifying different fluorescent reagents;

cell selection settings, number of cells to be analyzed per well, and range of size, shape, and intensity of cells to be analyzed.

8. The method of claim 1 wherein the feature data include one or more of:

size, shape, intensity, location, area, perimeter squared area, height width ratio, total fluorescence intensity, and average fluorescence intensity.

9. The method of claim 8 wherein the step of collecting well summary data includes calculating one or more of:

size, shape, intensity, location, area, perimeter squared area, height width ratio, total fluorescence intensity, and average fluorescent fluorescence intensity.