WO2004003129A2 - Treatment of tissue specimens - Google Patents

Treatment of tissue specimens Download PDF

Info

Publication number
WO2004003129A2
WO2004003129A2 PCT/GB2003/002570 GB0302570W WO2004003129A2 WO 2004003129 A2 WO2004003129 A2 WO 2004003129A2 GB 0302570 W GB0302570 W GB 0302570W WO 2004003129 A2 WO2004003129 A2 WO 2004003129A2
Authority
WO
WIPO (PCT)
Prior art keywords
analysis
tissues
specimens
research
distribution
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/GB2003/002570
Other languages
French (fr)
Other versions
WO2004003129A3 (en
Inventor
James Alexander Sharpe
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Medical Research Council
Original Assignee
Medical Research Council
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from GB0214846A external-priority patent/GB0214846D0/en
Priority claimed from GBGB0227649.1A external-priority patent/GB0227649D0/en
Priority to US10/518,307 priority Critical patent/US7677197B2/en
Priority to DK03740729T priority patent/DK1516183T3/en
Priority to AU2003280409A priority patent/AU2003280409B2/en
Priority to EP03740729A priority patent/EP1516183B1/en
Application filed by Medical Research Council filed Critical Medical Research Council
Priority to JP2004516916A priority patent/JP4129456B2/en
Priority to DE60313170T priority patent/DE60313170T2/en
Priority to CA2489096A priority patent/CA2489096C/en
Publication of WO2004003129A2 publication Critical patent/WO2004003129A2/en
Publication of WO2004003129A3 publication Critical patent/WO2004003129A3/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • G01N1/31—Apparatus therefor
    • 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
    • G01N15/1468—Optical investigation techniques, e.g. flow cytometry with spatial resolution of the texture or inner structure of the particle
    • 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
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/47—Scattering, i.e. diffuse reflection
    • G01N21/4795—Scattering, i.e. diffuse reflection spatially resolved investigating of object in scattering medium
    • 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
    • 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
    • G01N15/1468—Optical investigation techniques, e.g. flow cytometry with spatial resolution of the texture or inner structure of the particle
    • G01N2015/1472—Optical investigation techniques, e.g. flow cytometry with spatial resolution of the texture or inner structure of the particle with colour
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/02—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations
    • G01N35/04—Details of the conveyor system
    • G01N2035/0474—Details of actuating means for conveyors or pipettes
    • G01N2035/0477—Magnetic
    • 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

Definitions

  • TITLE TREATMENT OF TISSUE SPECIMENS
  • This invention relates to the treatment of tissue specimens, particularly (but not exclusively) prior to imaging of the specimens by optical projection tomography (OPT), and to uses of optical projection tomography methods and apparatus.
  • OPT optical projection tomography
  • Tissue specimens are normally treated by immersion in one or more liquids, before the specimens are imaged in OPT apparatus, and the invention was devised as a way of facilitating this pre-treatment of specimens.
  • apparatus for treating tissue specimens by immersion in a liquid comprising a first structure providing a chamber for holding the liquid, and a second structure including holding means for releasably holding the specimens, the first and second structures being relatively moveable in a direction having a vertical component between a first position in which the holding means are relatively close to the chamber and in which the second structure closes the top of the chamber to enable the specimens to be immersed in the liquid whilst the latter is protected from the environment, and a second position in which the holding means are relatively distant from the chamber to enable the specimens to be loaded onto or unloaded from the holding means.
  • the first structure is stationary and the second structure is shiftable vertically with respect to the first structure.
  • the holding means may include magnets to enable specimens, each provided with a metal mount, to be detachably retained on the second structure by magnetic attraction.
  • the chamber is preferably in the form of an annular trough in which case the holding means may hold the specimens so that the latter depend from the holding means, conveniently at angularly spaced positions around a circle such that the specimens are lowered into the trough as the second structure is lowered to its first position.
  • the second structure preferably includes a lid which acts to close the chamber in the first position and the underside of which carries the holding means. Lid closure helps to prevent evaporation of volatile treatment liquids.
  • the second structure may be rotatably moveable around a central vertical axis, enabling specimens to be loaded onto and unloaded from the second structure at a chosen position alongside the apparatus, either by a robotic arm or a human hand.
  • the apparatus may have the facility to change the liquid when in the first position, enabling the specimens to be treated by different liquids in a succession of treatment stages, whilst retaining the chamber closed.
  • the apparatus may have a pump to fill and empty the chamber with a succession of chosen liquids which, in the case of tissue specimens, may act to wash or otherwise treat the specimens prior to the specimens being imaged by means of optical projection tomography.
  • a method of treating tissue specimens by immersion in a liquid in a chamber comprising loading the specimens onto a holder so that the specimens depend from the holder and are disposed above the liquid in the chamber, effecting relative movement between the chamber and the specimens in one direction to cause immersion of the specimens in the liquid whilst maintaining the chamber closed and protected from the environment during immersion, effecting relative movement between the chamber and the specimens in the opposite direction to bring the specimens out of the liquid, and unloading the treated specimens from the holder.
  • the specimens may be treated by different liquids in a plurality of treatment stages which are preferably carried out by successive emptying and filling of the chamber with the different liquids, whilst the specimens remain in the chamber and whilst the chamber remains closed and protected from the environment.
  • analyses and procedures of the present invention include:
  • Analysis of the distribution of transgenic gene activity within biological tissues Analysis of the distribution of cell activities within biological tissues, including:
  • 5,5'-dibromo-4,4'-dichloro-indigo (or other halogenated indigo compounds) formazan or other coloured precipitates generated through the catalytic activity of enzymes including: b-galactosidase, alkaline phosphatase or other coloured precipitates formed upon catalytic conversion of staining substrates, including: Fast Red, Vector Red
  • any fluorescent substances such as: Alexa dyes, FITC, rhodamine,
  • GFP green fluorescent protein
  • invertebrates nematode worms vertebrates all types of fish including teleosts, such as zebrafish, and chondrycthes including sharks
  • amphibians including the genus Xenopus and axolotls
  • reptiles birds including chickens and quails
  • all mammals including all rodents, dogs, cats and all primates, including human
  • embryonic tissues for any purpose, including: research into any stem cell population research into developmental biology research into the causes of abnormal embryo development, including human syndromes autopsies of human terminated pregnancies (both spontaneous and induced terminations)
  • Analysis of any tissues for the purpose of genomics research including: the analysis of any tissues for the purpose of genomics research, including: the analysis of transgenic, knock-in, knock-down or knock-out organisms the analysis or discovery of the expression (or activity) of genes including their spatial distribution, and their levels of expression the analysis of discovery of abnormalities in the structure or morphology of tissues, as a result of interference due to wilful experimentation (such as genetic or physical modifications including a chemical or biochemical genomics approach), and/or spontaneous abnormalities (such as naturally- occurring mutations)
  • Analysis of any tissue for pharmaceutical research including: the analysis of pharmaceutical substances (such as drugs, molecules, proteins, antibodies), including their spatial distribution within the tissue, and their concentrations the analysis or discovery of abnormalities in the structure or morphology of tissues.
  • pharmaceutical substances such as drugs, molecules, proteins, antibodies
  • tissues for medical research including: research into the genetics, development, physiology, structure and function of animal tissues analysis of diseased tissue to further our understanding of all types of diseases including: congenital diseases acquired diseases including: infectious neoplastic vascular inflammatory traumatic metabolic endocrine degenerative drug-related iatrogenic or idiopathic diseases
  • Analysis of tissues for medical diagnosis, treatment or monitoring including: the diagnosis of cancer patients including: searching for cancerous cells and tissues within biopsies searching for abnormal structure or morphology of tissues within biopsies the analysis of all biopsies including the analysis of: lymph nodes polyps liver biopsies kidney biopsies prostate biopsies muscle biopsies brain tissue the analysis of tissue removed in the process of extracting a tumour from a patient including: determining whether all the tumour has been removed determining the type of tumour, and the type of cancer.
  • samples for use in the present invention may be prepared as described in the earlier patent applications and/or employing conventional pathological and histological techniques and procedures well known to persons skilled in the art.
  • Figure 1 is an isometric view of apparatus according to the invention and in an open condition
  • Figure 2 is an isometric view of the apparatus in a closed condition
  • Figure 3 is a fragmentary view, to an enlarged scale, of part of the apparatus in an open condition, showing specimens held by the apparatus,
  • Figure 4 is a cross-sectional view of the apparatus
  • Figure 5 shows the apparatus in combination with a robotic arm
  • Figure 6 is an enlarged view of part of Figure 5
  • Figure 7 shows the robotic arm positioning a specimen onto the rotary stage of an OPT scanner.
  • the apparatus comprises a fixed structure 1 having a cylindrical outer casing 2 in the top of which is positioned an annular trough 3 which is open at the top.
  • a moveable structure 4 having a central hub or spindle 5 on the top of which is mounted a disc-like lid 6 the underside of which carries a number of angularly spaced and downwardly depending cylindrical magnets 7.
  • the hub or spindle 5 is rotatably mounted in the fixed structure for rotation of the moveable structure about a central vertical axis indicated at 8 in Figure 4. Also, the hub or spindle 5 is capable of vertical translational movement with respect to the fixed structure 1 along the vertical axis 8.
  • the trough 3 is capable of being filled with liquids, and the structure 1 includes containers for holding these liquids and pumps for filling and emptying the trough, as indicated at 10 in Figure 4.
  • the magnets 7 are used to hold, in a detachable manner, tissue specimens 12 each of which has been prepared with a metal mount 13 at one end of the specimen. This allows each metal 13 mount to depend from one of the magnets 7, with the specimen 12 depending downwardly from the mount 13.
  • the magnets 7 are raised clear of the top of the trough 3 so that the specimens 12 can be attached to or removed from the magnets 7.
  • the lid 6 engages the top of the casing 2 and the specimens 12 are immersed in a liquid 14 in the trough 3.
  • the robotic arm 15 shown in Figures 6 and 7 is used to load untreated specimens into the apparatus and also to transfer treated specimens from the apparatus to the rotary stage 16 of an OPT scanner where the specimens are imaged.
  • the robotic arm 15 is used to load specimens into the apparatus, each specimen being attached to the lower end of a corresponding magnet 7 by virtue of the magnetic attraction between the magnet 7 and the metal mount 13 at one end of the specimen 12, the moveable structure 4 being in the open position and being indexed in a rotational sense as the specimens are loaded.
  • the moveable structure When loaded with specimens, the moveable structure is moved to its lowered or closed position, thereby immersing the specimens 12 in the liquid 14 which has been pumped into the trough 3. In this closed position, the lid 6 engages the upper rim of the outer casing 2 so that the liquid 14 is closed to the air, thereby allowing the use of a volatile liquid, unlike known apparatus which uses conveyor belts for transferring specimens through a liquid.
  • the first liquid is drained from the trough 3 and a second liquid pumped thereinto, without the need for the trough 3 to be opened to the air.
  • the attachment of the metal mounts 13 to the magnets 7 retains the specimens 12 in their hanging positions so that the specimens do not engage the bottom of the trough 3, which could damage them.
  • any number of treatment stages can be carried out in this manner, the liquid being changed without the need to move the specimens and employing only a small volume of each treatment liquid.
  • the moveable structure 4 After treatment of the specimens 12, the moveable structure 4 is raised to its upper position and the treated specimens 12 are then transferred to the rotary stage 16 by means of the robotic arm 15, the structure 4 being rotationally indexed to enable the robotic arm 15 to unload each specimen 12 in turn and to transfer the treated specimen 5 to the rotary stage 16.
  • liquids for treating the specimens are fixatives (such as paraformaldehyde or formalin), alcohols (in particular methanol and ethanol) and organic solvents for clearing the specimens (in particular benzyl alcohol and benzyl benzoate).
  • fixatives such as paraformaldehyde or formalin
  • alcohols in particular methanol and ethanol
  • organic solvents for clearing the specimens in particular benzyl alcohol and benzyl benzoate.
  • Analysis of the distribution of transgenic gene activity within biological tissues Analysis of the distribution of cell activities within biological tissues, including:
  • 5,5'-dibromo-4,4'-dichloro-indigo (or other halogenated indigo compounds) formazan or other coloured precipitates generated through the catalytic activity of enzymes including: b-galactosidase, alkaline phosphatase or other coloured precipitates formed upon catalytic conversion of staining substrates, including: Fast Red, Vector Red And including any light-emitting substances,
  • any fluorescent substances such as: Alexa dyes, FITC, rhodamine,
  • GFP green fluorescent protein
  • amphibians including the genus Xenopus and axolotls
  • reptiles birds including chickens and quails
  • all mammals including all rodents, dogs, cats and all primates, including human
  • embryonic tissues for any purpose, including: research into any stem cell population research into developmental biology research into the causes of abnormal embryo development, including human syndromes autopsies of human terminated pregnancies (both spontaneous and induced terminations)
  • any tissues for the purpose of genomics research including: the analysis of transgenic, knock-in, knock-down or knock-out organisms the analysis or discovery of the expression (or activity) of genes including their spatial distribution, and their levels of expression the analysis of discovery of abnormalities in the structure or morphology of tissues, as a result of interference due to wilful experimentation (such as genetic or physical modifications including a chemical or biochemical genomics approach), and/or spontaneous abnormalities (such as naturally-occurring mutations)
  • Analysis of any tissue for pharmaceutical research including: the analysis of pharmaceutical substances (such as drugs, molecules, proteins, antibodies), including their spatial distribution within the tissue, and their concentrations the analysis or discovery of abnormalities in the structure or morphology of tissues.
  • pharmaceutical substances such as drugs, molecules, proteins, antibodies
  • tissues for medical research including: research into the genetics, development, physiology, structure and function of animal tissues analysis of diseased tissue to further our understanding of all types of diseases including: congenital diseases acquired diseases including: infectious neoplastic vascular inflammatory traumatic metabolic endocrine degenerative drug-related iatrogenic or idiopathic diseases
  • Analysis of tissues for medical diagnosis, treatment or monitoring including: the diagnosis of cancer patients including: searching for cancerous cells and tissues within biopsies searching for abnormal structure or morphology of tissues within biopsies the analysis of all biopsies including the analysis of: lymph nodes polyps liver biopsies kidney biopsies prostate biopsies muscle biopsies brain tissue the analysis of tissue removed in the process of extracting a tumour from a patient including: determining whether all the tumour has been removed determming the type of tumour, and the type of cancer.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Immunology (AREA)
  • Optics & Photonics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Dispersion Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Sampling And Sample Adjustment (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

Apparatus for treating tissue specimens comprises a stationary outer casing (2) in the top of which is positioned an annular trough (3) for holding a liquid. Rotatably mounted in the outer casing (2) is a central hub (5) supporting a disc-like lid (6) the underside of which carries a magnet (7). In use, the magnets hold, in a detachable manner, tissue specimens (12) and the trough (3) is filled with a liquid for treating the specimens. The hub (5) is capable of vertical translational movement with respect to the casing (2) to enable the specimens to be lowered into and subsequently lifted out of the trough (3). A method of treating tissue specimens is also provided.

Description

TITLE: TREATMENT OF TISSUE SPECIMENS
Field of the Invention
This invention relates to the treatment of tissue specimens, particularly (but not exclusively) prior to imaging of the specimens by optical projection tomography (OPT), and to uses of optical projection tomography methods and apparatus.
Background to the Invention
Tissue specimens are normally treated by immersion in one or more liquids, before the specimens are imaged in OPT apparatus, and the invention was devised as a way of facilitating this pre-treatment of specimens.
Summary of the Invention
According to one aspect of the invention there is provided apparatus for treating tissue specimens by immersion in a liquid, the apparatus comprising a first structure providing a chamber for holding the liquid, and a second structure including holding means for releasably holding the specimens, the first and second structures being relatively moveable in a direction having a vertical component between a first position in which the holding means are relatively close to the chamber and in which the second structure closes the top of the chamber to enable the specimens to be immersed in the liquid whilst the latter is protected from the environment, and a second position in which the holding means are relatively distant from the chamber to enable the specimens to be loaded onto or unloaded from the holding means.
Preferably, the first structure is stationary and the second structure is shiftable vertically with respect to the first structure. The holding means may include magnets to enable specimens, each provided with a metal mount, to be detachably retained on the second structure by magnetic attraction.
The chamber is preferably in the form of an annular trough in which case the holding means may hold the specimens so that the latter depend from the holding means, conveniently at angularly spaced positions around a circle such that the specimens are lowered into the trough as the second structure is lowered to its first position. In this case, the second structure preferably includes a lid which acts to close the chamber in the first position and the underside of which carries the holding means. Lid closure helps to prevent evaporation of volatile treatment liquids.
The second structure may be rotatably moveable around a central vertical axis, enabling specimens to be loaded onto and unloaded from the second structure at a chosen position alongside the apparatus, either by a robotic arm or a human hand.
The apparatus may have the facility to change the liquid when in the first position, enabling the specimens to be treated by different liquids in a succession of treatment stages, whilst retaining the chamber closed. For example, the apparatus may have a pump to fill and empty the chamber with a succession of chosen liquids which, in the case of tissue specimens, may act to wash or otherwise treat the specimens prior to the specimens being imaged by means of optical projection tomography.
According to another aspect of the invention there is provided a method of treating tissue specimens by immersion in a liquid in a chamber, the method comprising loading the specimens onto a holder so that the specimens depend from the holder and are disposed above the liquid in the chamber, effecting relative movement between the chamber and the specimens in one direction to cause immersion of the specimens in the liquid whilst maintaining the chamber closed and protected from the environment during immersion, effecting relative movement between the chamber and the specimens in the opposite direction to bring the specimens out of the liquid, and unloading the treated specimens from the holder.
The specimens may be treated by different liquids in a plurality of treatment stages which are preferably carried out by successive emptying and filling of the chamber with the different liquids, whilst the specimens remain in the chamber and whilst the chamber remains closed and protected from the environment.
According to another aspect of the present invention, there is provided a method of performing any one or more of the analyses or procedures listed hereunder comprising use of a method or apparatus of any of the aspects set out above.
There is also provided use of a method or apparatus as described in any of the aspects as set out above in any one or more of the analyses or procedures listed hereunder.
According to the present invention, the analyses and procedures of the present invention include:
Analysis of the structure of biological tissues.
Analysis of the function of biological tissues.
Analysis of the shapes of biological tissues.
Analysis of the distribution of cell types within biological tissues.
Analysis of the distribution of gene activity within biological tissues, including the distribution of:
- RNA transcripts
- proteins
Analysis of the distribution of transgenic gene activity within biological tissues, Analysis of the distribution of cell activities within biological tissues, including:
- Cell cycle status including arrest
- Cell death - Cell proliferation
- Cell migration
Analysis of the distribution of physiological states within biological tissues. Analysis of the results of immunohistochemistry staining techniques. Analysis of the results of in-situ hybridisation staining techniques. Analysis of the distribution of molecular markers within biological tissues, including any coloured or light-absorbing substances, such as:
5,5'-dibromo-4,4'-dichloro-indigo (or other halogenated indigo compounds) formazan or other coloured precipitates generated through the catalytic activity of enzymes including: b-galactosidase, alkaline phosphatase or other coloured precipitates formed upon catalytic conversion of staining substrates, including: Fast Red, Vector Red
And including any light-emitting substances,
Therefore including any fluorescent substances, such as: Alexa dyes, FITC, rhodamine,
And including any luminescent substances, such as green fluorescent protein (GFP) or similar proteins,
And including any phosphorescent substances.
Analysis of tissues from all plant species. Analysis of any tissue for agricultural research, including: basic research into all aspects of plant biology (genetics, development, physiology, pathology etc.) analysis of tissues which have been genetically altered.
Analysis of tissues from all animal species, including: invertebrates nematode worms vertebrates all types of fish (including teleosts, such as zebrafish, and chondrycthes including sharks) amphibians (including the genus Xenopus and axolotls) reptiles birds (including chickens and quails) all mammals (including all rodents, dogs, cats and all primates, including human)
Analysis of embryonic tissues for any purpose, including: research into any stem cell population research into developmental biology research into the causes of abnormal embryo development, including human syndromes autopsies of human terminated pregnancies (both spontaneous and induced terminations)
Analysis of any tissues for the purpose of genomics research, including: the analysis of any tissues for the purpose of genomics research, including: the analysis of transgenic, knock-in, knock-down or knock-out organisms the analysis or discovery of the expression (or activity) of genes including their spatial distribution, and their levels of expression the analysis of discovery of abnormalities in the structure or morphology of tissues, as a result of interference due to wilful experimentation (such as genetic or physical modifications including a chemical or biochemical genomics approach), and/or spontaneous abnormalities (such as naturally- occurring mutations)
Analysis of any tissue for the purpose of neurobiology research, including: the analysis of the morphology of nerves the analysis of the pathways and connectivity of nerves the analysis of parts of, or whole, animal brains
Analysis of any tissue for pharmaceutical research, including: the analysis of pharmaceutical substances (such as drugs, molecules, proteins, antibodies), including their spatial distribution within the tissue, and their concentrations the analysis or discovery of abnormalities in the structure or morphology of tissues.
Analysis of tissues for medical research, including: research into the genetics, development, physiology, structure and function of animal tissues analysis of diseased tissue to further our understanding of all types of diseases including: congenital diseases acquired diseases including: infectious neoplastic vascular inflammatory traumatic metabolic endocrine degenerative drug-related iatrogenic or idiopathic diseases
Analysis of tissues for medical diagnosis, treatment or monitoring, including: the diagnosis of cancer patients including: searching for cancerous cells and tissues within biopsies searching for abnormal structure or morphology of tissues within biopsies the analysis of all biopsies including the analysis of: lymph nodes polyps liver biopsies kidney biopsies prostate biopsies muscle biopsies brain tissue the analysis of tissue removed in the process of extracting a tumour from a patient including: determining whether all the tumour has been removed determining the type of tumour, and the type of cancer.
According to the present invention, samples for use in the present invention may be prepared as described in the earlier patent applications and/or employing conventional pathological and histological techniques and procedures well known to persons skilled in the art.
For example, in-situ hybridisation (particularly useful for detecting RNAs):Hammond K L, Hanson I M, Brown A G, Lettice L A, Hill R E "Mammalian and Drosophila dachsund genes are related to the Ski proto-oncongene and are expressed in eye and limb" . Mech Dev. 1998 Jun;74(l-2): 121-31. Immunohistochemistry (particularly useful for detecting proteins and other molecules): Sharpe J, Ahlgren U, Perry P, Hill B, Ross A, Hecksher-Sorensen J, Baldock R, Davidson D. "Optical projection tomography as a tool for 3D microscopy and gene expression studies" Science. 2002 Apr 19;296(5567):541-5.
It will be appreciated that modification may be made to the invention without departing from the scope of the invention.
Brief Description of the Drawings
The invention will now be further described, by way of example, with reference to the accompanying drawings, in which :-
Figure 1 is an isometric view of apparatus according to the invention and in an open condition,
Figure 2 is an isometric view of the apparatus in a closed condition,
Figure 3 is a fragmentary view, to an enlarged scale, of part of the apparatus in an open condition, showing specimens held by the apparatus,
Figure 4 is a cross-sectional view of the apparatus,
Figure 5 shows the apparatus in combination with a robotic arm,
Figure 6 is an enlarged view of part of Figure 5, and
Figure 7 shows the robotic arm positioning a specimen onto the rotary stage of an OPT scanner. Detailed Description
The apparatus comprises a fixed structure 1 having a cylindrical outer casing 2 in the top of which is positioned an annular trough 3 which is open at the top. Mounted in the fixed structure is a moveable structure 4 having a central hub or spindle 5 on the top of which is mounted a disc-like lid 6 the underside of which carries a number of angularly spaced and downwardly depending cylindrical magnets 7. The hub or spindle 5 is rotatably mounted in the fixed structure for rotation of the moveable structure about a central vertical axis indicated at 8 in Figure 4. Also, the hub or spindle 5 is capable of vertical translational movement with respect to the fixed structure 1 along the vertical axis 8.
Within the casing 2 are located motors and gearing for driving the structure 4 both in rotation and translational movement, as indicated at 9 in Figure 4. The trough 3 is capable of being filled with liquids, and the structure 1 includes containers for holding these liquids and pumps for filling and emptying the trough, as indicated at 10 in Figure 4.
The magnets 7 are used to hold, in a detachable manner, tissue specimens 12 each of which has been prepared with a metal mount 13 at one end of the specimen. This allows each metal 13 mount to depend from one of the magnets 7, with the specimen 12 depending downwardly from the mount 13. When the apparatus is in the open condition (Figure 1), the magnets 7 are raised clear of the top of the trough 3 so that the specimens 12 can be attached to or removed from the magnets 7. When the apparatus is in the closed position (Figure 2), the lid 6 engages the top of the casing 2 and the specimens 12 are immersed in a liquid 14 in the trough 3.
The robotic arm 15 shown in Figures 6 and 7 is used to load untreated specimens into the apparatus and also to transfer treated specimens from the apparatus to the rotary stage 16 of an OPT scanner where the specimens are imaged. In use, the robotic arm 15 is used to load specimens into the apparatus, each specimen being attached to the lower end of a corresponding magnet 7 by virtue of the magnetic attraction between the magnet 7 and the metal mount 13 at one end of the specimen 12, the moveable structure 4 being in the open position and being indexed in a rotational sense as the specimens are loaded. When loaded with specimens, the moveable structure is moved to its lowered or closed position, thereby immersing the specimens 12 in the liquid 14 which has been pumped into the trough 3. In this closed position, the lid 6 engages the upper rim of the outer casing 2 so that the liquid 14 is closed to the air, thereby allowing the use of a volatile liquid, unlike known apparatus which uses conveyor belts for transferring specimens through a liquid.
If it is required to treat the specimens by a succession of liquids, the first liquid is drained from the trough 3 and a second liquid pumped thereinto, without the need for the trough 3 to be opened to the air. Moreover, the attachment of the metal mounts 13 to the magnets 7 retains the specimens 12 in their hanging positions so that the specimens do not engage the bottom of the trough 3, which could damage them.
Any number of treatment stages can be carried out in this manner, the liquid being changed without the need to move the specimens and employing only a small volume of each treatment liquid.
After treatment of the specimens 12, the moveable structure 4 is raised to its upper position and the treated specimens 12 are then transferred to the rotary stage 16 by means of the robotic arm 15, the structure 4 being rotationally indexed to enable the robotic arm 15 to unload each specimen 12 in turn and to transfer the treated specimen 5 to the rotary stage 16.
Examples of liquids for treating the specimens are fixatives (such as paraformaldehyde or formalin), alcohols (in particular methanol and ethanol) and organic solvents for clearing the specimens (in particular benzyl alcohol and benzyl benzoate). The apparatus and methods can be used in various analyses and procedures, as set out below:
Analysis of the structure of biological tissues.
Analysis of the function of biological tissues.
Analysis of the shapes of biological tissues.
Analysis of the distribution of cell types within biological tissues.
Analysis of the distribution of gene activity within biological tissues, including the distribution of:
- RNA transcripts
- proteins
Analysis of the distribution of transgenic gene activity within biological tissues, Analysis of the distribution of cell activities within biological tissues, including:
- Cell cycle status including arrest
- Cell death
- Cell proliferation
- Cell migration
Analysis of the distribution of physiological states within biological tissues. Analysis of the results of immunohistochemistry staining techniques. Analysis of the results of in-situ hybridisation staining techniques. Analysis of the distribution of molecular markers within biological tissues, including any coloured or light-absorbing substances, such as:
5,5'-dibromo-4,4'-dichloro-indigo (or other halogenated indigo compounds) formazan or other coloured precipitates generated through the catalytic activity of enzymes including: b-galactosidase, alkaline phosphatase or other coloured precipitates formed upon catalytic conversion of staining substrates, including: Fast Red, Vector Red And including any light-emitting substances,
Therefore including any fluorescent substances, such as: Alexa dyes, FITC, rhodamine,
And including any luminescent substances, such as green fluorescent protein (GFP) or similar proteins,
And including any phosphorescent substances.
Analysis of tissues from all plant species. Analysis of any tissue for agricultural research, including: basic research into all aspects of plant biology (genetics, development, physiology, pathology etc.) analysis of tissues which have been genetically altered.
Analysis of tissues from all animal species, including: invertebrates nematode worms vertebrates all types of fish
(including teleosts, such as zebrafish, and chondrycthes including sharks) amphibians (including the genus Xenopus and axolotls) reptiles birds (including chickens and quails) all mammals (including all rodents, dogs, cats and all primates, including human)
Analysis of embryonic tissues for any purpose, including: research into any stem cell population research into developmental biology research into the causes of abnormal embryo development, including human syndromes autopsies of human terminated pregnancies (both spontaneous and induced terminations)
Analysis of any tissues for the purpose of genomics research, including: the analysis of transgenic, knock-in, knock-down or knock-out organisms the analysis or discovery of the expression (or activity) of genes including their spatial distribution, and their levels of expression the analysis of discovery of abnormalities in the structure or morphology of tissues, as a result of interference due to wilful experimentation (such as genetic or physical modifications including a chemical or biochemical genomics approach), and/or spontaneous abnormalities (such as naturally-occurring mutations)
Analysis of any tissue for the purpose of neurobiology research, including: the analysis of the morphology of nerves the analysis of the pathways and connectivity of nerves the analysis of parts of, or whole, animal brains
Analysis of any tissue for pharmaceutical research, including: the analysis of pharmaceutical substances (such as drugs, molecules, proteins, antibodies), including their spatial distribution within the tissue, and their concentrations the analysis or discovery of abnormalities in the structure or morphology of tissues.
Analysis of tissues for medical research, including: research into the genetics, development, physiology, structure and function of animal tissues analysis of diseased tissue to further our understanding of all types of diseases including: congenital diseases acquired diseases including: infectious neoplastic vascular inflammatory traumatic metabolic endocrine degenerative drug-related iatrogenic or idiopathic diseases
Analysis of tissues for medical diagnosis, treatment or monitoring, including: the diagnosis of cancer patients including: searching for cancerous cells and tissues within biopsies searching for abnormal structure or morphology of tissues within biopsies the analysis of all biopsies including the analysis of: lymph nodes polyps liver biopsies kidney biopsies prostate biopsies muscle biopsies brain tissue the analysis of tissue removed in the process of extracting a tumour from a patient including: determining whether all the tumour has been removed determming the type of tumour, and the type of cancer.
It will be appreciated that modification may be made to the invention without departing from the scope of the invention.

Claims

1. Apparatus for treating tissue specimens by immersion in a liquid, the apparatus comprising a first structure providing a chamber for holding the liquid, and a second structure including holding means for releasably holding the specimens, the first and second structures being relatively moveable in a direction having a vertical component between a first position in which the holding means are relatively close to the chamber and in which the second structure closes the top of the chamber to enable the specimens to be immersed in the liquid whilst the latter is protected from the environment, and a second position in which the holding means are relatively distant from the chamber to enable the specimens to be loaded onto or unloaded from the holding means.
2. Apparatus according to claim 1, wherein the first structure is stationary and the second structure is shiftable vertically with respect to the first structure.
3. Apparatus according to claim 1 or 2, wherein the holding means include magnets to enable specimens, each provided with a metal mount, to be detachably retained on the second structure by magnetic attraction.
4. Apparatus according to any of the preceding claims, wherein the chamber is in the form of an annular trough.
5. Apparatus according to claim 4, wherein the holding means hold the specimens so that the latter depend from the holding means at angularly spaced positions around a circle such that the specimens are lowered into the trough as the second structure is lowered to its first position.
6. Apparatus according to claim 4 or 5, wherein the second structure includes a lid which acts to close the chamber in the first position and the underside of which carries the holding means.
7. Apparatus according to any of the preceding claims, wherein the second structure is rotatably moveable around a central vertical axis, enabling specimens to be loaded onto and unloaded from the second structure at a chosen position alongside the apparatus, either by a robotic arm or a human hand.
8. Apparatus according to any of the preceding claims, wherein the apparatus has the facility to change the liquid when in the first position, enabling the specimens to be treated by different liquids in a succession of treatment stages, whilst retaining the chamber closed.
9. A method of treating tissue specimens by immersion in a liquid in a chamber, the method comprising loading the specimens onto a holder so that the specimens depend from the holder and are disposed above the liquid in the chamber, effecting relative movement between the chamber and the specimens in one direction to cause immersion of the specimens in the liquid whilst maintaining the chamber closed and protected from the environment during immersion, effecting relative movement between the chamber and the specimens in the opposite direction to bring the specimens out of the liquid, and unloading the treated specimens from the holder.
10. A method according to claim 9, wherein the specimens are treated by different liquids in a plurality of treatment stages.
11. A method according to claim 10, wherein the treatment stages are carried out by successive emptying and filling of the chamber with the different liquids, whilst the specimens remain in the chamber and whilst the chamber remains closed and protected from the environment.
12. A method of performing any one or more of the analyses or procedures listed hereunder comprising use of a method or apparatus according to any of claims 1 to 11:
Analysis of the structure of biological tissues.
Analysis of the function of biological tissues.
Analysis of the shapes of biological tissues.
Analysis of the distribution of cell types within biological tissues.
Analysis of the distribution of gene activity within biological tissues, including the distribution of:
- RNA transcripts
- proteins
Analysis of the distribution of transgenic gene activity within biological tissues, Analysis of the distribution of cell activities within biological tissues, including:
- Cell cycle status including arrest
- Cell death
- Cell proliferation
- Cell migration
Analysis of the distribution of physiological states within biological tissues. Analysis of the results of immunohistochemistry staining techniques. Analysis of the results of in-situ hybridisation staining techniques. Analysis of the distribution of molecular markers within biological tissues, including any coloured or light-absorbing substances, such as:
5,5'-dibromo-4,4'-dichloro-indigo (or other halogenated indigo compounds) formazan or other coloured precipitates generated through the catalytic activity of enzymes including: b-galactosidase, alkaline phosphatase or other coloured precipitates formed upon catalytic conversion of staining substrates, including: Fast Red, Vector Red
And including any light-emitting substances,
Therefore including any fluorescent substances, such as: Alexa dyes, FITC, rhodamine,
And including any luminescent substances, such as green fluorescent protein (GFP) or similar proteins,
And including any phosphorescent substances.
Analysis of tissues from all plant species. Analysis of any tissue for agricultural research, including: basic research into all aspects of plant biology (genetics, development, physiology, pathology etc.) analysis of tissues which have been genetically altered.
Analysis of tissues from all animal species, including: invertebrates nematode worms vertebrates all types of fish (including teleosts, such as zebrafish, and chondrycthes including sharks) amphibians (including the genus Xenopus and axolotls) reptiles birds (including chickens and quails) all mammals (including all rodents, dogs, cats and all primates, including human)
Analysis of embryonic tissues for any purpose, including: research into any stem cell population research into developmental biology research into the causes of abnormal embryo development, including human syndromes autopsies of human terminated pregnancies (both spontaneous and induced terminations)
Analysis of any tissues for the purpose of genomics research, including: the analysis of any tissues for the purpose of genomics research, including: the analysis of transgenic, knock-in, knock-down or knock-out organisms the analysis or discovery of the expression (or activity) of genes including their spatial distribution, and their levels of expression the analysis of discovery of abnormalities in the structure or morphology of tissues, as a result of interference due to wilful experimentation (such as genetic or physical modifications including a chemical or biochemical genomics approach), and/or spontaneous abnormalities (such as naturally- occurring mutations)
Analysis of any tissue for the purpose of neurobiology research, including: the analysis of the morphology of nerves the analysis of the pathways and connectivity of nerves the analysis of parts of, or whole, animal brains
Analysis of any tissue for pharmaceutical research, including: the analysis of pharmaceutical substances (such as drugs, molecules, proteins, antibodies), including their spatial distribution within the tissue, and their concentrations the analysis or discovery of abnormalities in the structure or morphology of tissues.
Analysis of tissues for medical research, including: research into the genetics, development, physiology, structure and function of animal tissues analysis of diseased tissue to further our understanding of all types of diseases including: congenital diseases acquired diseases including: infectious neoplastic vascular inflammatory traumatic metabolic endocrine degenerative drug-related iatrogenic or idiopathic diseases
Analysis of tissues for medical diagnosis, treatment or monitoring, including: the diagnosis of cancer patients including: searching for cancerous cells and tissues within biopsies searching for abnormal structure or morphology of tissues within biopsies the analysis of all biopsies including the analysis of: lymph nodes polyps liver biopsies kidney biopsies prostate biopsies muscle biopsies brain tissue the analysis of tissue removed in the process of extracting a tumour from a patient including: determining whether all the tumour has been removed determining the type of tumour, and the type of cancer.
13. Use of a method or apparatus as described in any of claims 1 to 11 in any one or more of the analyses or procedures listed hereunder:
Analysis of the structure of biological tissues.
Analysis of the function of biological tissues.
Analysis of the shapes of biological tissues.
Analysis of the distribution of cell types within biological tissues.
Analysis of the distribution of gene activity within biological tissues, including the distribution of:
- RNA transcripts
- proteins
Analysis of the distribution of transgenic gene activity within biological tissues, Analysis of the distribution of cell activities within biological tissues, including:
- Cell cycle status including arrest
- Cell death
- Cell proliferation
- Cell migration
Analysis of the distribution of physiological states within biological tissues. Analysis of the results of immunohistochemistry staining techniques. Analysis of the results of in-situ hybridisation staining techniques. Analysis of the distribution of molecular markers within biological tissues, including any coloured or light-absorbing substances, such as: 5,5'-dibromo-4,4'-dichloro-indigo (or other halogenated indigo compounds) formazan or other coloured precipitates generated through the catalytic activity of enzymes including: b-galactosidase, alkaline phosphatase or other coloured precipitates formed upon catalytic conversion of staining substrates, including: Fast Red, Vector Red
And including any light-emitting substances,
Therefore including any fluorescent substances, such as: Alexa dyes, FITC, rhodamine,
And including any luminescent substances, such as green fluorescent protein (GFP) or similar proteins,
And including any phosphorescent substances.
Analysis of tissues from all plant species. Analysis of any tissue for agricultural research, including: basic research into all aspects of plant biology (genetics, development, physiology, pathology etc.) analysis of tissues which have been genetically altered.
Analysis of tissues from all animal species, including: invertebrates nematode worms vertebrates all types of fish (including teleosts, such as zebrafish, and chondrycthes including sharks) amphibians (including the genus Xenopus and axolotls) reptiles birds (including chickens and quails) all mammals (including all rodents, dogs, cats and all primates, including human) Analysis of embryonic tissues for any purpose, including: research into any stem cell population research into developmental biology research into the causes of abnormal embryo development, including human syndromes autopsies of human terminated pregnancies (both spontaneous and induced terminations)
Analysis of any tissues for the purpose of genomics research, including: the analysis of any tissues for the purpose of genomics research, including: the analysis of transgenic, knock-in, knock-down or knock-out organisms the analysis or discovery of the expression (or activity) of genes including their spatial distribution, and their levels of expression the analysis of discovery of abnormalities in the structure or morphology of tissues, as a result of interference due to wilful experimentation (such as genetic or physical modifications including a chemical or biochemical genomics approach), and/or spontaneous abnormalities (such as naturally- occurring mutations)
Analysis of any tissue for the purpose of neurobiology research, including: the analysis of the morphology of nerves the analysis of the pathways and connectivity of nerves the analysis of parts of, or whole, animal brains
Analysis of any tissue for pharmaceutical research, including: the analysis of pharmaceutical substances (such as drugs, molecules, proteins, antibodies), including their spatial distribution within the tissue, and their concentrations the analysis or discovery of abnormalities in the structure or morphology of tissues.
Analysis of tissues for medical research, including: research into the genetics, development, physiology, structure and function of animal tissues analysis of diseased tissue to further our understanding of all types of diseases including: congenital diseases acquired diseases including: infectious neoplastic vascular inflammatory traumatic metabolic endocrine degenerative drug-related iatrogenic or idiopathic diseases
Analysis of tissues for medical diagnosis, treatment or monitoring, including: the diagnosis of cancer patients including: searching for cancerous cells and tissues within biopsies searching for abnormal strucmre or morphology of tissues within biopsies the analysis of all biopsies including the analysis of: lymph nodes polyps liver biopsies kidney biopsies prostate biopsies muscle biopsies brain tissue the analysis of tissue removed in the process of extracting a tumour from a patient including: determining whether all the mmour has been removed determining the type of tumour, and the type of cancer.
PCT/GB2003/002570 2002-06-27 2003-06-13 Treatment of tissue specimens Ceased WO2004003129A2 (en)

Priority Applications (7)

Application Number Priority Date Filing Date Title
CA2489096A CA2489096C (en) 2002-06-27 2003-06-13 Treatment of tissue specimens
DE60313170T DE60313170T2 (en) 2002-06-27 2003-06-13 TREATMENT OF TISSUE SAMPLES
DK03740729T DK1516183T3 (en) 2002-06-27 2003-06-13 Treatment of tissue samples
AU2003280409A AU2003280409B2 (en) 2002-06-27 2003-06-13 Treatment of tissue specimens
EP03740729A EP1516183B1 (en) 2002-06-27 2003-06-13 Treatment of tissue specimens
US10/518,307 US7677197B2 (en) 2002-06-27 2003-06-13 Treatment of tissue specimens
JP2004516916A JP4129456B2 (en) 2002-06-27 2003-06-13 Tissue sample processing

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
GB0214846.8 2002-06-27
GB0214846A GB0214846D0 (en) 2002-06-27 2002-06-27 Treatment of tissue specimens
GBGB0227649.1A GB0227649D0 (en) 2002-11-27 2002-11-27 Uses of optical projection tomography methods and apparatus
GB0227649.1 2002-11-27

Publications (2)

Publication Number Publication Date
WO2004003129A2 true WO2004003129A2 (en) 2004-01-08
WO2004003129A3 WO2004003129A3 (en) 2004-02-26

Family

ID=30001980

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB2003/002570 Ceased WO2004003129A2 (en) 2002-06-27 2003-06-13 Treatment of tissue specimens

Country Status (11)

Country Link
US (1) US7677197B2 (en)
EP (1) EP1516183B1 (en)
JP (1) JP4129456B2 (en)
AT (1) ATE359507T1 (en)
AU (1) AU2003280409B2 (en)
CA (1) CA2489096C (en)
DE (1) DE60313170T2 (en)
DK (1) DK1516183T3 (en)
ES (1) ES2285147T3 (en)
PT (1) PT1516183E (en)
WO (1) WO2004003129A2 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004048970A1 (en) * 2002-11-27 2004-06-10 Medical Research Council Uses of optical projection tomography methods and apparatus

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB0503986D0 (en) * 2005-02-26 2005-04-06 Secr Defence Reaction apparatus
EP3485976A1 (en) * 2017-11-21 2019-05-22 Koninklijke Philips N.V. Method of handling a biopsy sample
CN108801752B (en) * 2018-08-02 2023-11-28 佛山科学技术学院 A sample loading device and a sample driving device

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2583379A (en) * 1949-03-16 1952-01-22 Technicon Company Automatic immersion apparatus
US3062225A (en) * 1960-04-11 1962-11-06 Corode Electrochemie S P R L Machine for immersing work-pieces in containers
US4202289A (en) * 1978-07-10 1980-05-13 Bils Robert F Specimen processing apparatus for microscopy
GB8628371D0 (en) * 1986-11-27 1986-12-31 Shandon Southern Prod Tissue specimen treatment apparatus
US6355104B1 (en) * 1998-11-20 2002-03-12 Louis S. Polster Article treating apparatus and method and control system therefor
US6562136B1 (en) * 2000-09-08 2003-05-13 Surmodics, Inc. Coating apparatus and method
GB0112392D0 (en) * 2001-05-22 2001-07-11 Medical Res Council Optical imaging appartus and associated specimen support means

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004048970A1 (en) * 2002-11-27 2004-06-10 Medical Research Council Uses of optical projection tomography methods and apparatus

Also Published As

Publication number Publication date
DK1516183T3 (en) 2007-08-13
DE60313170D1 (en) 2007-05-24
ES2285147T3 (en) 2007-11-16
EP1516183A2 (en) 2005-03-23
AU2003280409A1 (en) 2004-01-19
AU2003280409A2 (en) 2004-01-19
CA2489096C (en) 2013-08-06
JP4129456B2 (en) 2008-08-06
WO2004003129A3 (en) 2004-02-26
US20060105332A1 (en) 2006-05-18
JP2005531005A (en) 2005-10-13
AU2003280409B2 (en) 2008-08-21
DE60313170T2 (en) 2007-08-23
PT1516183E (en) 2007-04-30
US7677197B2 (en) 2010-03-16
ATE359507T1 (en) 2007-05-15
EP1516183B1 (en) 2007-04-11
CA2489096A1 (en) 2004-01-08

Similar Documents

Publication Publication Date Title
US20210131925A1 (en) Method for making biological material transparent and use thereof
KR100743727B1 (en) Rapid tissue processor
US10444124B2 (en) Clarifying reagent for biological materials and use thereof
US20210131926A1 (en) Clearing reagent for biological material, and use thereof
US6793890B2 (en) Rapid tissue processor
AU2003287895B2 (en) Method and apparatus for pretreatment of biological samples
JP5058676B2 (en) Biological specimen preparation method
EP3732461B1 (en) Tissue sample preparation system
JP2014005231A (en) Method for making biological material transparent and kit for clearing treatment for making biological material transparent
KR20010023328A (en) Method and apparatus for automatically forming monolayers from particulate matter separated from fluid samples
Miscevic et al. Advances in zebrafish high content and high throughput technologies
EP2518468B1 (en) Method, processor and carrier for processing frozen slices of tissue of biospecimens
US20060105332A1 (en) Treatment of tissue specimens
Yu et al. Amphioxus whole-mount in situ hybridization
WO2004048970A1 (en) Uses of optical projection tomography methods and apparatus
Oisi et al. Analysis of embryonic gene expression patterns in the hagfish
French et al. Balancing essential and non-essential metal bioavailability during hatchery rearing of Greenshell mussel (Perna canaliculus) larvae
WO2020147035A1 (en) Automated liquid handling system and method for depositing biological samples for microscopic examination
Durán et al. Freeze substitution followed by low melting point wax embedding preserves histomorphology and allows protein and mRNA localization techniques

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NI NO NZ OM PH PL PT RO RU SC SD SE SG SK SL TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IT LU MC NL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
WWE Wipo information: entry into national phase

Ref document number: 2489096

Country of ref document: CA

WWE Wipo information: entry into national phase

Ref document number: 2003280409

Country of ref document: AU

WWE Wipo information: entry into national phase

Ref document number: 2003740729

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 2004516916

Country of ref document: JP

WWP Wipo information: published in national office

Ref document number: 2003740729

Country of ref document: EP

ENP Entry into the national phase

Ref document number: 2006105332

Country of ref document: US

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 10518307

Country of ref document: US

WWP Wipo information: published in national office

Ref document number: 10518307

Country of ref document: US

WWG Wipo information: grant in national office

Ref document number: 2003740729

Country of ref document: EP