EP1017982A1 - Dispositif permettant de sortir un echantillon d'un ensemble d'echantillons et outil coupant pour ledit dispositif - Google Patents
Dispositif permettant de sortir un echantillon d'un ensemble d'echantillons et outil coupant pour ledit dispositifInfo
- Publication number
- EP1017982A1 EP1017982A1 EP98946150A EP98946150A EP1017982A1 EP 1017982 A1 EP1017982 A1 EP 1017982A1 EP 98946150 A EP98946150 A EP 98946150A EP 98946150 A EP98946150 A EP 98946150A EP 1017982 A1 EP1017982 A1 EP 1017982A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- sample
- cutting tool
- array
- cutting
- samples
- 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.)
- Withdrawn
Links
- 238000000034 method Methods 0.000 claims description 13
- 239000000463 material Substances 0.000 claims description 12
- 239000003302 ferromagnetic material Substances 0.000 claims description 8
- 238000003491 array Methods 0.000 claims description 3
- 239000004033 plastic Substances 0.000 claims description 3
- 229920003023 plastic Polymers 0.000 claims description 3
- 238000000151 deposition Methods 0.000 claims description 2
- 239000011521 glass Substances 0.000 claims description 2
- 239000000696 magnetic material Substances 0.000 claims description 2
- 238000001454 recorded image Methods 0.000 claims description 2
- 238000005070 sampling Methods 0.000 abstract description 9
- 238000012546 transfer Methods 0.000 abstract description 5
- 239000000523 sample Substances 0.000 description 47
- 239000000499 gel Substances 0.000 description 18
- 239000012528 membrane Substances 0.000 description 8
- 238000012360 testing method Methods 0.000 description 8
- 238000004458 analytical method Methods 0.000 description 6
- 229920000642 polymer Polymers 0.000 description 4
- 239000004677 Nylon Substances 0.000 description 3
- 238000011005 laboratory method Methods 0.000 description 3
- 229920001778 nylon Polymers 0.000 description 3
- 229920006254 polymer film Polymers 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 description 2
- 239000004743 Polypropylene Substances 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 239000000853 adhesive Substances 0.000 description 2
- 230000001070 adhesive effect Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 229910052751 metal Inorganic materials 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 229920005597 polymer membrane Polymers 0.000 description 2
- -1 polypropylene Polymers 0.000 description 2
- 229920001155 polypropylene Polymers 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 239000011240 wet gel Substances 0.000 description 2
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 229920000544 Gore-Tex Polymers 0.000 description 1
- 239000000020 Nitrocellulose Substances 0.000 description 1
- 239000002033 PVDF binder Substances 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- NIXOWILDQLNWCW-UHFFFAOYSA-N acrylic acid group Chemical group C(C=C)(=O)O NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 125000003275 alpha amino acid group Chemical group 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 235000014510 cooky Nutrition 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 239000002783 friction material Substances 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 230000005291 magnetic effect Effects 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000000520 microinjection Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229920001220 nitrocellulos Polymers 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 229920001343 polytetrafluoroethylene Polymers 0.000 description 1
- 239000004810 polytetrafluoroethylene Substances 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 230000013777 protein digestion Effects 0.000 description 1
- 238000001742 protein purification Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- WFKWXMTUELFFGS-UHFFFAOYSA-N tungsten Chemical compound [W] WFKWXMTUELFFGS-UHFFFAOYSA-N 0.000 description 1
- 229910052721 tungsten Inorganic materials 0.000 description 1
- 239000010937 tungsten Substances 0.000 description 1
- 238000000539 two dimensional gel electrophoresis Methods 0.000 description 1
- 238000012800 visualization Methods 0.000 description 1
Definitions
- the present invention relates to a apparatus for removing one or more samples from an array of samples and to a cutting tool for use with that apparatus.
- the invention relates to an apparatus for excising and ejecting biomolecules from an array of biomolecule samples in a gel or solid support.
- WO 97/29355 discloses a process and an apparatus for effecting contactless transfer of special compounds into biological objects, typically cells, located on a polymer carrier film by laser microinjection.
- the laser is also used to cut out the object, by cutting around it.
- the laser is a U-V laser and the polymer film is a special U-V absorbing polymer film.
- the sample is then impelled by the laser beam to an adjacent adhesive collecting film.
- the collecting film has to be located between 1 and 10mm from the polymer film, preferably 1-3 mm. Locating the sample to be cut out is done by fluorescence microscopy which produces an image of the table on which the carrier film is located to enable the laser to be guided for injection and cutting.
- the present invention seeks to overcome or at least ameliorate some of the problems associated with the background art discussed above. Summary of the Invention
- the present invention relates to a method for excising at least one sample in an array of samples comprising:
- steps (a) to (d) depositing the at least one excised sample at the selected location.
- the steps (a) to (d) may be repeated or cycled so as to carry out a series of excisions of a number of different samples in the array.
- the samples may be separated into the array by known means such as electrophoresis in a polymer matrix.
- Samples may be transferred from the polymer matrix onto a solid support or membrane support.
- two dimensional electrophoresis separations in polyacrylamide are transferred to supports like PTFE, gortex, PVDF, nylon, nitrocellulose, polypropylene which are particularly suitable for supporting an array of samples for excision using the methods and apparatus of the present invention.
- Samples may also be excised without having to transfer them to a membrane.
- the present invention relates to an apparatus for excising at least one sample from an array of samples comprising:-
- the sampling device may further include: e) a table means for supporting the array of samples; f) display means for displaying the electronic image of the array on a screen or the like; g) means for selecting a sample on the array for sampling by the cutting tool; h) means for moving the cutting tool in the plane of the array.
- the array of samples will be present as a non-ordered array of spots on dry polymer membranes or wet gels.
- the sample will preferably be of a biological nature and may include proteins, peptides, polysaccharides, lipids and nucleic acid molecules or complex molecules like glycoproteins, for example.
- the means for recording an electronic image of at least a part of the array of samples may be a digital camera which makes a digital photograph of the samples.
- the electronic image may be generated from a scan of the samples stained or illuminated or otherwise marked with a visible or fluorescent marker to allow them to be visualised.
- An ink jet dispensing unit such as is disclosed in applicants co-pending International patent application No PCT/AU98/00265, the entire contents of which are incorporated herein by reference, could be used for marking the spots/samples.
- the coordinates of the samples are recorded. These coordinates are then transformed into robotic language and the computer used to control a cutting tool whereupon the cutting tool can be directed to a selected sample. The position of all the samples would be known from their co-ordinates on the grid, and so excision is possible regardless of whether or not the samples are still visible.
- the cutting tool moves to a selected coordinate of a well of a microtitre plate or the like and the sample is ejected
- an image file relating to a number of arrays of samples is stored on the computer. An image of a particular array is displayed on the computer and spots are selected from the array to be sampled using the computer monitor and a mouse. Once a mouse is clicked on a particular spot the cutting tool will automatically move to that spot, cut the spot from the array, pick up the spot and transport it to the selected well, or the like.
- the array of samples is in a plane, the x-y plane, and the table means is movable in both the x and y directions so that the spot to be sampled is placed underneath the cutting tool.
- the table means fixed and the cutting tool movable.
- a cutting tool for use in the apparatus and method of the present invention comprises: a cutting head defining a central bore adapted to cut and retain a sample of material; and a plunger disposed in the bore defining a rod which is disposed in and movable along the bore, the plunger being either formed of a ferromagnetic material or having a portion of ferro-magnetic material attached thereto; a solenoid disposed around the plunger or electromagnetic material, wherein operation of the solenoid causes the plunger to move to eject the spot from the cutting head.
- the punch may be circular. In one embodiment the cutting device is pneumatically operated.
- the return means may also be a magnet.
- the return means may also be a spring.
- the cutting tip is removable and disposable.
- Figure 1 is a front view of a first sampling device embodying the present invention:
- FIG. 2 is an enlarged front view of a cutting tool assembly which forms part of the sampling device of Figure 1;
- Figure 3 is a side view of the cutting tool assembly shown in Figure 2;
- Figure 4 and 5 are detailed views of the cutting head of the cutting tool assembly, where 4 shows a sample pick-up and 5 a sample eject;
- Figures 6a to 6h show the sequence of operation of the cutting tool assembly showing in sequence the cutting of a sample from an array of samples and the placing of that cut sample in a test tube;
- Figure 7 shows a schematic of the electronic recording and sample excision robot aspects of a second apparatus according to the present invention.
- Figure 8a shows a second embodiment of an excision tool
- Figure 8b shows a third embodiment of an excision tool.
- Figure 1 shows a sampling device embodying the present invention generally indicated at 10.
- the sampling device includes a table 12 and a overhead beam 14 which is spaced and supported above the table by two columns 16 and 18.
- a cutting tool assembly 20 is located which is described in more detail below.
- the table 12 is mounted so as to be moveable in the x and y directions in a generally horizontal plane. By moving the table in the x and y directions any part of the table may be located under the cutting tool assembly 20.
- Motors and control means are provided for moving the table. The specific means for moving the table in the x and y directions is not essential to the present invention.
- FIG. 2 and Figure 3 show an enlarged front view and side view respectively of the cutting tool assembly.
- the cutting tool assembly 20 is supported on a guide means 22 which maintains the components of the assembly in their correct orientation.
- a top cylinder plate 24 which is horizontally oriented.
- a bottom cylinder plate 26 Spaced below and parallel to, the top cylinder plate there is a bottom cylinder plate 26.
- a cylinder 28 is disposed and extends between the two plates.
- the cylinder forms part of a pneumatic piston and cylinder arrangement which includes a piston rod 30 shown in phantom in figure 3, disposed in the cylinder 28.
- the depending free end 32 of the piston rod is fixed to a nylon block which defines a punch carriage 34 disposed directly below the bottom cylinder plate 26. Movement of the piston rod 30 in the cylinder 28 in the vertical (z axis) direction causes the punch carriage 34 to move in the vertical direction guided by guide rods. Movement of the piston in the cylinder is pneumatically controlled.
- the punch carriage 34 defines a cylindrical aperture or bore 35 which extends up from the lower face 34A of the punch carriage, towards the piston rod 30 and the upper end 34B of the punch carriage.
- a cutting head assembly generally indicated at 36 is disposed inside that aperture 35 and is shown in more detail in Figures 4 and 5.
- a solenoid 38 is disposed in the interior of the aperture 35.
- a generally cylindrical nylon block 40 is fixed in the centre of the aperture 35 with its longitudinal axis directly in line with the piston rod 30.
- a punch holder 42 is disposed inside a central bore of the block 40.
- the punch holder defines a central bore 44 in which a steel plunger 46 is located.
- the plunger 46 is generally cylindrical and defines a large diameter body portion 48 from one lower end 48A of which there depends a narrower cylindrical rod 50.
- a spring 52 is disposed between the opposite or upper end 48B of the plunger and the upper end of the central bore 44.
- a punch 54 is fixed to the lower end of the punch holder/cutting head assembly.
- a generally cylindrical bore 56 extends through the centre of the punch 54.
- the cylindrical rod 50 of the plunger is disposed in that bore.
- the rod 50 is coated with a low friction material such as TEFLONTM or ACETYLTM both as a barrier to prevent residue from previously picked up samples from contaminating subsequently taken samples and to reduce friction between the rod 50 and bore 56.
- the lower end of the punch defines a generally circular cutting blade or tip 58, similar to a cookie cutter.
- Figures 6a to 6h show the cutting and transfer sequence of the sampling device of the present invention in sequence in more detail.
- the cutting head is disposed above a particular spot which is to be cut out from, for example, a solid (i.e. non-gel) s ipport which may be an immobilisation membrane.
- the punch carriage 34 is located at the upper end of its travel and the end 50A of the plunger projects beyond the end of the punch 54.
- the punch carriage is operated by the pneumatic piston and cylinder assembly to push the cutting head downwards until the end 50A of the plunger touches the spot/sample to be cut. This contact secures the spot 94 and ensures it does not move.
- the cutting head is moved downwards towards the test tvibe so that the plunger is positioned inside the test tube.
- the plunger is then pushed downwards by activating the solenoid which causes the plunger to move downwards and eject the sample 94.
- the cutting head will then automatically move to cut the next sample.
- the procedure is slightly different when cutting a spot from a sheet of gel to allow for the fact that gel is easily squashed.
- the plunger is retracted. This prevents squashing of the gel which would occur if the end 50A projected below the punch and were forced onto the gel.
- the procedure for ejecting the gel spot is different also. After the cutting head has been positioned in the test tube lifting of the head is commenced fractionally (say lms) before the solenoid is activated to eject the gel spot, also to prevent squashing the spot.
- the device also includes a digital camera adapted to create an image of the gel or membrane on which the spots are located and store them in a computer.
- the computer is programmed with software which takes account of the distortions produced by the digital camera when imaging the array and produces a sufficiently accurate distortion free map of the array which accords with the mechanical frame used to control the cutting head assembly.
- the gel or membrane There is also no need for the gel or membrane to be exactly planar. Different programs are provided for gel and non-gel arrays to account for the slightly different procedures described above.
- the image of the array can be displayed on a computer monitor and a mouse moved by an operator to identify the correct spot to be sampled by the sampling device.
- the present invention allows a vary laborious job of a researcher individually cutting spots from a gel or support using a scalpel and allows the operation to be carried out automatically.
- FIG. 7 shows a schematic representation of a second embodiment of a robotic excision apparatus.
- the robotic excision apparatus 100 includes an image acquisition system 200 which includes a camera, an excision tool 400, and a computer 300.
- An array of samples is placed onto a silicon mat 105 which is housed inside an acrylic base plate 101 which is illuminated from underneath the sample with fluorescent light (for acrylamide) or from above with tungsten lamps or a camera flash (for membranes) 106.
- the image is transferred from the image acquisition means to the computer 300.
- the image is processed and imported into "click-on-a-spot" software. This process translates the image pixel coordinates into robot coordinates.
- the "click-on-a-spot" software is then used to drive the excision tool 400 to the selected component via an xy movable bar 102.
- the z movement of the excision tool 400 is via an excision tool support unit 107.
- the excision tool 400 which is described in more detail below, then cuts ovit and holds the selected sample and moves above a specific well of a microtitre plate into which the sample is to be placed.
- the excised sample is then deposited into the specific well of the microtitre plate 108.
- a first embodiment of the excision/cutting tool for use with the apparatus of Figure 7 is shown in Figure 8a.
- the cutting tool comprises a cylindrical body portion 400, which has an upper end 400 A and a lower end 400B.
- the body portion is a generally cylindrical tube defining a central bore 408 and can be made of metal or hard plastic or any suitable material.
- the lower end 400B of the tube is closed with a end portion which acts as a tip holder 404 which has a central cylindrical bore in which is mounted a tip 409.
- the tip 409 has an annular cross section and has a wider cylindrical portion which locates inside the bore 408 of the body and after it emerges below the tip holder then tapers generally conically to a narrower portion which acts as a cutting head 406.
- the tip, and tip holder are fixed relative to the body portion.
- the tip can be made of various materials including glass, metal or a plastic, such as polypropylene. However, it is preferable that the tip is translucent as this makes it possible to determine if acrylamide residue is caught inside the tip.
- the tip may be removable and disposable.
- the pin is not illustrated in Figure 8a but its upper end is fixed to an ejector magnet 402 and the lower end of the ejector pin should be at least 2mm higher than the orifice of the cutting head when the ejector pin is in its uppermost, retracted or "home" position.
- a solenoid 401 surrounds the ejector magnetic 402.
- a "spring" magnet 410 having a central bore along which the ejector pin is free to move is disposed between the ejector magnet 408 and the tip portion 404. The spring magnet 410 is oriented so that it repels the ejector magnet 402 upwards so that the ejector pin is normally retracted.
- Figure 8b shows an alternative arrangement in which a spring 403 is used to keep the ejector magnet 402 in the "up" position instead of a magnet.
- the solenoid 401 of the cutting tool body 400 is activated which drives the ejector magnet 402 down onto a spring mechanism 403. This forces the ejector pin 405 through the cutting head orifice 406 ejecting the sample into a microtitre plate.
- the solenoid is deactivated the ejector magnet 402 is forced back up into the solenoid body by the expansion of the spring 403.
- the cutting tip 416 shown in Figure 8b is disposable and is generally conical, with a gentle taper and an annular cross-section. It pushes or snap- fits onto a conical protrusion 418 depending from the body 400.
- the cutting tool should be of sufficient length so that the ejector pin is at least 2 mm higher than the orifice of the cutting head when the ejector magnet is in the home position
- the ejector pin should protrude from the cutting tool orifice by at least 1mm when the ejector magnet is forced down by the activation of the solenoid.
- the cutting tool rotates about its central longitudinal axis to facilitate cutting of the sample.
- the tool may be mounted on a screw thread extending in the z axis direction.
Landscapes
- Sampling And Sample Adjustment (AREA)
Abstract
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AUPO9438A AUPO943897A0 (en) | 1997-09-25 | 1997-09-25 | Sampling device |
AUPO943897 | 1997-09-25 | ||
AUPO361598 | 1998-05-20 | ||
AUPO361598 | 1998-05-20 | ||
PCT/AU1998/000813 WO1999015875A1 (fr) | 1997-09-25 | 1998-09-25 | Dispositif permettant de sortir un echantillon d'un ensemble d'echantillons et outil coupant pour ledit dispositif |
Publications (2)
Publication Number | Publication Date |
---|---|
EP1017982A1 true EP1017982A1 (fr) | 2000-07-12 |
EP1017982A4 EP1017982A4 (fr) | 2004-10-06 |
Family
ID=25645314
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP98946150A Withdrawn EP1017982A4 (fr) | 1997-09-25 | 1998-09-25 | Dispositif permettant de sortir un echantillon d'un ensemble d'echantillons et outil coupant pour ledit dispositif |
Country Status (1)
Country | Link |
---|---|
EP (1) | EP1017982A4 (fr) |
Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4354406A (en) * | 1980-01-14 | 1982-10-19 | L'oreal | Method of and device for forming openings in a gelatinous substance |
JPH03240482A (ja) * | 1990-02-16 | 1991-10-25 | Hitachi Electron Eng Co Ltd | 液体培地へのコロニーの移植方法 |
US5334353A (en) * | 1993-02-03 | 1994-08-02 | Blattner Frederick R | Micropipette device |
US5587062A (en) * | 1996-01-24 | 1996-12-24 | Shimadzu Corporation | Sample collecting apparatus by gel electrophoresis |
US5638170A (en) * | 1994-06-07 | 1997-06-10 | Hudson Control Group, Inc. | Punched carrier specimen sample processor |
WO1997032666A1 (fr) * | 1996-03-05 | 1997-09-12 | Rainin Instrument Co., Inc. | Pipette manuelle dotee d'un element de verrouillage en position initiale a retardement |
WO1998023950A1 (fr) * | 1996-11-29 | 1998-06-04 | Oxford Glycosciences (Uk) Ltd. | Gels, procedes et appareil d'identification et de caracterisation de biomolecules |
-
1998
- 1998-09-25 EP EP98946150A patent/EP1017982A4/fr not_active Withdrawn
Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4354406A (en) * | 1980-01-14 | 1982-10-19 | L'oreal | Method of and device for forming openings in a gelatinous substance |
JPH03240482A (ja) * | 1990-02-16 | 1991-10-25 | Hitachi Electron Eng Co Ltd | 液体培地へのコロニーの移植方法 |
US5334353A (en) * | 1993-02-03 | 1994-08-02 | Blattner Frederick R | Micropipette device |
US5638170A (en) * | 1994-06-07 | 1997-06-10 | Hudson Control Group, Inc. | Punched carrier specimen sample processor |
US5587062A (en) * | 1996-01-24 | 1996-12-24 | Shimadzu Corporation | Sample collecting apparatus by gel electrophoresis |
WO1997032666A1 (fr) * | 1996-03-05 | 1997-09-12 | Rainin Instrument Co., Inc. | Pipette manuelle dotee d'un element de verrouillage en position initiale a retardement |
WO1998023950A1 (fr) * | 1996-11-29 | 1998-06-04 | Oxford Glycosciences (Uk) Ltd. | Gels, procedes et appareil d'identification et de caracterisation de biomolecules |
Non-Patent Citations (2)
Title |
---|
PATENT ABSTRACTS OF JAPAN vol. 016, no. 025 (C-0903), 22 January 1992 (1992-01-22) & JP 03 240482 A (HITACHI ELECTRON ENG CO LTD), 25 October 1991 (1991-10-25) * |
See also references of WO9915875A1 * |
Also Published As
Publication number | Publication date |
---|---|
EP1017982A4 (fr) | 2004-10-06 |
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