Classifications

• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
  • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
  • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
  • G01N21/64—Fluorescence; Phosphorescence
  • G01N21/645—Specially adapted constructive features of fluorimeters
  • G01N21/6456—Spatial resolved fluorescence measurements; Imaging
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
  • G01N15/10—Investigating individual particles
  • G01N15/14—Optical investigation techniques, e.g. flow cytometry
  • G01N15/1429—Signal processing
  • G01N15/1433—Signal processing using image recognition
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
  • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
  • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
  • G01N21/64—Fluorescence; Phosphorescence
  • G01N21/645—Specially adapted constructive features of fluorimeters
  • G01N21/6452—Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
  • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
  • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
  • G01N21/64—Fluorescence; Phosphorescence
  • G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
• • G—PHYSICS
  • G06—COMPUTING OR CALCULATING; COUNTING
  • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
  • G06T7/00—Image analysis
  • G06T7/10—Segmentation; Edge detection
  • G06T7/11—Region-based segmentation
• • G—PHYSICS
  • G06—COMPUTING OR CALCULATING; COUNTING
  • G06V—IMAGE OR VIDEO RECOGNITION OR UNDERSTANDING
  • G06V20/00—Scenes; Scene-specific elements
  • G06V20/60—Type of objects
  • G06V20/69—Microscopic objects, e.g. biological cells or cellular parts
• • G—PHYSICS
  • G06—COMPUTING OR CALCULATING; COUNTING
  • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
  • G06T2207/00—Indexing scheme for image analysis or image enhancement
  • G06T2207/10—Image acquisition modality
  • G06T2207/10056—Microscopic image
• • G—PHYSICS
  • G06—COMPUTING OR CALCULATING; COUNTING
  • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
  • G06T2207/00—Indexing scheme for image analysis or image enhancement
  • G06T2207/10—Image acquisition modality
  • G06T2207/10064—Fluorescence image
• • G—PHYSICS
  • G06—COMPUTING OR CALCULATING; COUNTING
  • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
  • G06T2207/00—Indexing scheme for image analysis or image enhancement
  • G06T2207/30—Subject of image; Context of image processing
  • G06T2207/30004—Biomedical image processing
  • G06T2207/30024—Cell structures in vitro; Tissue sections in vitro

Definitions

• a method of imaging one or more biological objects using imaging apparatus capable of capturing an image across an imaging area, said method including: a) placing said one or more biological objects in an environment; b) providing in said environment, outside of said one or more biological objects, a contrast enhancing agent which provides contrast in an image between said one or more biological objects and said environment; and c) recording an image of said one or more biological objects and said environment using said imaging apparatus, whereby a spatial definition for said one or more biological objects is derivable using contrast in said image which is provided by said contrast enhancing agent.
• the invention also provides, in a further aspect, apparatus arranged to perform the above method.
• multi-well plate 180 which contains a two-dimensional array of sample wells 182.
• the optical elements of Figure 2 are depicted in cross-section and the multi-well plate in perspective.
• the projection of the line of illumination onto multi-well plate 180 is depicted by line 184 and is also understood to be perpendicular to the plane of Figure 2.
• multi-well plate 180 may be moved in two dimensions (X, Y) parallel to the dimensions of the array by means not shown.
• the cells 236 are distributed in a material located outside of the cells 236 which contains a predetermined concentration of a contrast enhancing agent (CEA) which is distributed approximately homogeneously throughout the material.
• CEA contrast enhancing agent
• the material is a medium 237 which is non-toxic to the cells 236, which supports the cells 236 by providing nutrients and conditions which maintain a health of the cells 236 and which helps the cell culture to grow.
• the cells 236 form a layer across the base plate 234 which has a thickness of approximately one cell, hi further embodiments, the layer may have a thickness which is greater than one cell.
• the imaging system additionally records, in addition to the image recorded in accordance with previous embodiments, a second image 260 of the cells and the environment across the imaging area in a second channel of the imaging system.
• the second CEA provides a contrast between relatively light areas 262 of the second image which corresponds to cell nuclei of the cells and relatively dark areas 264 corresponding to areas surrounding the cell nuclei which include the cell cytoplasm and the medium.
• These two recorded images are co-registered such that the pixels of each image are aligned.
• This is saved in a file known as an image stack.
• An image stack is a collection of images that logically belong together for analysis purposes. In most cases, a given image stack will contain images that share a common acquisition.
• the segmented object areas are next assigned S3 as cells, and regions outside of these segmented areas are assigned as the medium.
• relatively dark object areas of the image are separated out and assigned as the cells.
• relatively light object areas of the thresholded image are separated out and assigned as the cells.

Landscapes

• Health & Medical Sciences (AREA)
• Engineering & Computer Science (AREA)
• Physics & Mathematics (AREA)
• Life Sciences & Earth Sciences (AREA)
• General Physics & Mathematics (AREA)
• General Health & Medical Sciences (AREA)
• Chemical & Material Sciences (AREA)
• Pathology (AREA)
• Immunology (AREA)
• Analytical Chemistry (AREA)
• Biochemistry (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Theoretical Computer Science (AREA)
• Biomedical Technology (AREA)
• Molecular Biology (AREA)
• Computer Vision & Pattern Recognition (AREA)
• Multimedia (AREA)
• Signal Processing (AREA)
• Dispersion Chemistry (AREA)
• Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
• Apparatus For Radiation Diagnosis (AREA)

Priority Applications (3)

Applications Claiming Priority (2)

Publications (2)

Family

ID=34073478

Family Applications (1)

Country Status (5)

Cited By (4)

Families Citing this family (6)

Family Cites Families (9)

• 2004
  • 2004-12-10 GB GBGB0427050.0A patent/GB0427050D0/en not_active Ceased
• 2005
  • 2005-12-09 US US11/721,185 patent/US7881509B2/en not_active Expired - Fee Related
  • 2005-12-09 EP EP05820416A patent/EP1831668A2/en not_active Withdrawn
  • 2005-12-09 JP JP2007544985A patent/JP2008523376A/ja active Pending
  • 2005-12-09 WO PCT/GB2005/004747 patent/WO2006061640A2/en not_active Ceased

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