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WO1999039176A1 - Systemes de manutention mecanique destines a la microdissection par piegeage laser - Google Patents

Systemes de manutention mecanique destines a la microdissection par piegeage laser Download PDF

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Publication number
WO1999039176A1
WO1999039176A1 PCT/US1999/001987 US9901987W WO9939176A1 WO 1999039176 A1 WO1999039176 A1 WO 1999039176A1 US 9901987 W US9901987 W US 9901987W WO 9939176 A1 WO9939176 A1 WO 9939176A1
Authority
WO
WIPO (PCT)
Prior art keywords
cellular material
transporting substrate
tissue sample
identified cellular
selectively activated
Prior art date
Application number
PCT/US1999/001987
Other languages
English (en)
Inventor
Seth R. Goldstein
Robert F. Bonner
Paul D. Smith
John Peterson
Thomas Pohida
Original Assignee
The Government Of The United States Of America, Represented By The Secretary Of The Department Of Health And Human Services
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
Application filed by The Government Of The United States Of America, Represented By The Secretary Of The Department Of Health And Human Services filed Critical The Government Of The United States Of America, Represented By The Secretary Of The Department Of Health And Human Services
Priority to AU24850/99A priority Critical patent/AU749378B2/en
Priority to EP99904451A priority patent/EP1053461A1/fr
Priority to JP2000529587A priority patent/JP4344474B2/ja
Priority to US09/601,559 priority patent/US6720191B1/en
Priority to CA002319589A priority patent/CA2319589C/fr
Publication of WO1999039176A1 publication Critical patent/WO1999039176A1/fr

Links

Classifications

    • 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
    • G01N35/00009—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor provided with a sample supporting tape, e.g. with absorbent zones
    • 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
    • G01N1/2813—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on
    • 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
    • G01N1/2813—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on
    • G01N2001/282—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on with mapping; Identification of areas; Spatial correlated pattern
    • 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
    • G01N1/2813—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on
    • G01N2001/2833—Collecting samples on a sticky, tacky, adhesive surface
    • 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
    • G01N1/2813—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on
    • G01N2001/2833—Collecting samples on a sticky, tacky, adhesive surface
    • G01N2001/284—Collecting samples on a sticky, tacky, adhesive surface using local activation of adhesive, i.e. Laser Capture Microdissection

Definitions

  • This invention relates to laser capture microdissection (LCM) in which direct extraction of cellular material from a tissue sample occurs to a transfer surface.
  • LCM laser capture microdissection
  • the disclosure herein relates to isolating selected tissue samples to a film matrix in a format where the isolated sample segments can be conveniently collected for subsequent analysis .
  • LCM Laser Capture Microdissection
  • the substrate is then activated, typically be being pulsed with laser beam.
  • the light energy is absorbed by the plastic film which melts in a 2 small region.
  • the activatable coating then flows onto and around microscopic tissue components thereby causing the film on cooling to be firmly adherent to the identified cellular material of tissue sample.
  • Other target areas on the same slide can similarly treated.
  • the identified cellular material is dispersely distributed on the transporting substrate.
  • the selected series of spots is then removed from the film by placing the film in a suitable reagent (e.g., proteinase K which digests the structural proteins of the tissue) which frees the molecules of DNA or RNA to be subsequently analyzed (using, e.g., PCR and gel electrophoresis) .
  • a suitable reagent e.g., proteinase K which digests the structural proteins of the tissue
  • the transferred tissue spots and underlying attached film can be manually excised with a scalpel or scissors, or punched out with a precision punch/die directly into a cuvette.
  • the spots in the film are randomly located roughly in a position corresponding to their location on the sample.
  • Precise location is required so that the extracted portions of the sample can be precisely cut out.
  • the target tissue is completely recovered with minimal contamination from surrounding areas of film with low density nonspecific tissue adhesion.
  • precise computer control of the storage position and storage of coordinates may be practical. However, this computer control may loose the reliability of visual observation of the transfer process and may be complex and expensive. Further, computer location could fail to be accurate in many cases such as when the tape is distorted when it is lifted off the slide.
  • Nonspecific transfer to the film of un-targeted tissue in the region surrounding the identified cellular material is another problem. This non-specific transfer becomes increasingly important as spot size of targeted tissue is reduced. Ideally, no tissue outside the targeted region illuminated by the laser should be attached to the film upon its removal from the slide. Attachment of any undesired tissue would cause sample contamination with the desired tissue. As smaller and smaller target spots are used, the amount of stray tissue which can be tolerated becomes proportionally smaller.
  • a problem then arises as to how to handle the transfer surface. Specifically, the positioning of a small spot of activatable transfer surface on the tissue at the target site, picking the small activated transfer surface off the tissue, and placing the isolated targeted cells in a 4 cuvette, or storing them in a specifically identifiable manner without contamination by un-targeted tissue elements.
  • a method and apparatus of gathering by LCM identified cellular material from random locations on a tissue sample to designated locations on a transporting substrate enables convenient further processing.
  • a transporting substrate has an identified mapped location for receiving identified cellular material.
  • At least a segment of a selectively activatable coating is placed on the side of the transporting substrate in apposition to the tissue sample at the mapped location.
  • the transporting substrate and sample are relatively moved to place the selectively activated coating at the mapped location in apposition to identified cellular material of the tissue sample which is to be extracted.
  • the selectively activating coating is activated and impressed or impressed and activated to form an adhesive region on the transporting substrate for adhering to the identified cellular material.
  • identified cellular material adheres to the transporting substrate at the mapped location.
  • individual small pieces or coatings of selectively activated material are placed on a transporting film.
  • a selectively activatable coating is placed at the center of discrete pieces of film and exposed toward the sample.
  • Each different piece of film is separately activated at its coated center by the laser. Apposition to the tissue during or after laser action occurs by using a pressure plate.
  • a continuous strip including a transparent substrate is used to hold the film, such as a continuous reel of tape with equally spaced, centrally located pieces of selectively activatable coatings. Thereafter, the continuous strip of transparent substrate is 5 incrementally advanced so that center of the activatable coating is in the center of the microscope field. Adjacent bar codes or other optical encoded identifiers could serve to identify the individual transferred samples.
  • Sample collection can include a pressure plate actuated to hold the transporting substrate in contact with the tissue in the center of the microscope field before ' or after laser heating. After laser heating and attachment to the selected material from the sample, the pressure plate is raised, and the transporting substrate with the local activated coating with adherent tissue separated from the tissue sample.
  • This process is sequentially repeated so that the transporting substrate is again advanced, the next piece of transporting substrate (clean) is put into the center of the field, pressed onto the tissue surface, and laser activated.
  • This repetition not only advances the transporting substrate but also applies forces to reproducibly lift the targeted (adherent) tissue off the specimen slide.
  • the microscope stage and transporting substrate will be translated (in x & y) so that the next tissue target is in the center of the microscope field.
  • the substrate can be locally removed from the remainder substrate tape by punching or cutting off or peeling off the local regions of activated coating. This will leave attached identified cellular material localized to small pieces of transporting substrate. These small pieces of transporting substrate can be placed into a specific chamber or vessel for further processing and molecular analysis.
  • the individual pieces of selectively activatable coating are mounted at the ends of arrays of deflectable struts attached to and projecting from a central common support, such as a wheel or a comb like structure.
  • a central common support such as a wheel or a comb like structure.
  • Each strut with its selectively activatable coating at the end acts as the transporting substrate and is sequentially indexed through the center of the microscope 6 field.
  • the ends of the struts serve as pressure plates, contacting the tissue with the activatable coating at the ends of the struts.
  • the selectively activatable coat can be actuated when the strut is deflected, e.g., by a solenoid, (one spoke at a time, similar to a daisy wheel printer) .
  • the force on the strut is removed, allowing the strut to lift off the slide with the identified cellular material adhered to it. Thereafter, the strut array indexes the next strut with its piece of film into place, the stage can be moved to the next tissue target and the next strut is placed in contact with the new target.
  • Another variant of this approach is to use a comb shaped array (i.e, target film spots are in a linear array attached to a linear carriage by parallel spokes perpendicular to the array axis rather than the radial spokes of a "daisy wheel") .
  • a minute spot of film is placed at the end of each comb finger.
  • a small solenoid deflects the finger so its end, which acts as a pressure plate, contacts the tissue in the center of the field. After laser heating, the solenoid force is removed, the finger returns to its undeflected position (lifting off the slide the tissue adhered to the film) , and the comb is linearly indexed so a different finger/piece of film is in the center of the field and a different tissue target can be moved in place. Marks on the base of the comb could identify the samples. This technique allows extremely small and precise sample extraction.
  • the strut at the attached identified cellular material can be broken off, and this broken off portion deposited into a capsule for collection and further processing.
  • a piece of continuous transporting substrate is provided with a continuous central and narrow selectively activatable coating.
  • This piece of continuous film is put in contact with the tissue at the narrow selectively activatable coating by a narrow (in the direction of the film travel) pressure plate to define the (small) area of contact.
  • This approach has the advantage that it enables a small contact area of adhesive film on the tissue without: a) complications due to handling and alignment between the film and the target area, b) requiring specialized developments in the film above and beyond its basic adhesive properties, and c) requiring development of a specialized tape cassette e.g. with a built in pressure plate etc.
  • FIG. 1 is a perspective view of a mechanism for the practice of LCM including a supply reel, a laser activation system, a pressure plate for pushing the activated coating against a sample, and a cutting mechanism for generating discrete film sections with selected portions of the specimen contained thereon;
  • Fig. 2A-2E are a side elevation cartoon series of the sequential removal of selected sample from a slide mounted specimen in which: 8
  • Fig. 2A is illustrates the transporting film with coating attached to a selected specimen at a slide with the pressure plate withdrawn from contact with the slide;
  • Fig. 2B illustrates the apparatus of Fig. 2A with the tape being advanced to withdraw the selected portion of the sample from the slide ;
  • Fig. 2C illustrates the tape completely advanced for receipt of the next sequential sample
  • Fig. 2D illustrates the tape clamped as advance prior to collection of the next sample section
  • Fig. 2E illustrates the pressure plate pushing the activatable coating against the next sequentially selected portion of the sample
  • Fig. 3 is a schematic of tape being disposed parallel to the elongate axis of an underlying side with the pressure plate having transverse excursion relative to the slide to enable X and Y excursion of the sample collection point of the apparatus;
  • Fig. 4 is a plan view of a first embodiment of transporting tape utilized with this invention in which the film from the supply reel includes separate and discrete evenly spaced spots of selectively activatable coatings for the collection of sample;
  • Fig. 5 is a plan view of a second embodiment of the transporting tape here utilizing a separate central strip of selectively activatable coating for impressment to the sample;
  • Fig. 6 is a plan view of a tape having detachable sections with activatable coatings on the detachable sections;
  • Fig. 7 is a side elevation perspective illustrating a tape having a covering layer and illustrating the placement of marker coding on either the transporting substrate or the covering layer for subsequent retrieval of the tape;
  • Fig. 8 is a perspective view of a circular spoke apparatus having pads with selectively activatable coatings for the collection of selected sample from a specimen;
  • Fig. 9 is a perspective view of a linear spoke array of tines having an overall comb like configuration in which 9 sample is collected by individually actuating successive comb tines to collect designated sample ;
  • Fig. 10 is a detail at a spoke with the spoke acting through a transporting substrate to impress an activatable coating onto tissue sample with the activatable coating placed at the end of the spoke;
  • Fig. 11 illustrates sample being extracted from a tape to a collection vial
  • Fig. 12 illustrates sample being collected from a spoke to a collection vial
  • Fig. 13A illustrates an apparatus in which a sample is visualized with the results of visualization being placed into memory
  • Fig. 13B illustrates the slide of Fig. 13A with schematically indicated selected cellular material locations
  • Fig. 13C illustrates the slide of Fig. 13A transferred to apparatus activated to remotely undertake laser capture micro dissection utilizing the memory prepared by the apparatus of Fig. 13A;
  • Figs. 14A-14E are an illustration of activation of the coating in non-contact laser capture microdissection;
  • Fig. 14A shows the selectively activated coating on one side of the transporting substrate recessed from the target tissue by a gap;
  • Fig. 14B illustrates schematically laser energy being directed through the tissue sample to the selectively activated coating to activate that coating and cause the activated coating to expand downward to the sample at the identified cellular material;
  • Fig. 14C illustrates the transporting substrate and selectively activated coating after adhesion of the cellular material and contraction of the selectively activated coating to the transporting substrate;
  • Fig. 14D illustrates the transporting substrate of Fig. 14C with a new and different slide and tissue sample placed under the substrate
  • Fig. 14E illustrates the transporting substrate having samples from two differing slides, the collection here 10 illustrated showing an effective concentration selected cellular material, even though the selected material was contained on two separate samples .
  • Stepper motor 14 drives supply reel 16 dispensing coated tape 18.
  • Tape stop mechanism 24 includes stop bar
  • tape cut off knife 30 enables tape with contained sample S to be cut into small strips for further processing.
  • Slide 32 contains sample S, which sample includes desired sample sections 34 mixed irregularly with tissue 36 of sample S. It is the purpose of this disclosure to isolate with precision desired sample sections 34 from sample S. In order from such isolation to occur, visualization of sample S must occur.
  • microscope M having an inverted optical path looking up from under slide 32 is shown schematically by eye E.
  • slide 32 is mounted to a stage which is transportable in x-direction 38 and y-direction 40.
  • coated tape 18 is activated by laser light source 42. After or during such activation, the activated coating on coated tape 18 is contacted to sample S at desired sample sections 34. This contacting or juxtaposition between coated tape 18 and desired sample sections 34 occurs by pressure plate P bearing down on uncoated side 17 of coated tape 18 to cause the tape to move into apposition with sample S at desired sample sections 34. Adhesion of desired sample sections 34 occurs to coated side
  • tape tensioning apparatus 44 It may be desirable to maintain constant tension between supply reel 16 on dispensed coated tape 18. Accordingly, there is provided tape tensioning apparatus 44. It will be understood that this apparatus is optional. Once coated tape 18 has adhered to desired sample sections 34, the tape is moved. Under the normal circumstance, the tape will have sufficient adhesion to desired sample sections 34 to dissect the sample section from the remainder of the tissue of sample S. Referring to Figs. 2A - 2F, a schematic cartoon series of this apparatus is illustrated. In Fig. 2A, supply reel 16 is shown having coated tape 18 with coated side 19 in contact with sample S on slide 32. Because of previous activation, coated tape 18 is now adhered to desired sample sections 34 of sample S. As can be seen in the view of Figs 2A and 2F, pressure plate P is withdrawn.
  • stepper motor 14 causes a measured amount of coated tape 18 to be withdrawn from supply reel 16. When the limit of this measured amount is reached, stepper motor 14 freezes, and coated tape 18 is ready to be cut by tape cut off knife 30. Naturally, with stepper motor 14 precisely controlling dispensing of coated tape 18, the placement of desired sample sections 34 relative to the overall length cut off at tape cut off knife 30 occurs.
  • the transporting substrate has an identified mapped location for receiving identified cellular material.
  • tape segment 50 has been cut and removed. Further, tape stop mechanism 24 has been activated. As a consequence, the end of coated tape 18 is a fixed distance from P and is disposed for the collection of the next 12 visualized sample when pressure plate P deflects the tape onto the slide.
  • FIG. 2D it will be seen that slide 32 and consequently sample S have been moved to a new position. In this new position, another desired sample section 34 has been centered for pick up by movement of slide 32 in x- direction 38 and y-direction 40.
  • pressure plate P causes juxtaposition of coated tape 18 at coated side 19.
  • Stepper motor 14 is no longer frozen in order to provide the amount of slack necessary to deflect the tape onto slide 32. Again activation of coated side 19 by laser light source 42 through laser light focusing lens 46 occurs.
  • Fig. 2F the cycle illustrated is ready for repeat; it being seen that Fig. 2F illustrates the same cycle state as that previous observed in Fig. 2A.
  • a segment of a selectively activated coating is placed on the side of the transporting substrate in apposition to the tissue sample what amounts to a mapped location. Further, and during the gathering of desired sample sections 34, the transporting substrate and sample are relatively moved to place the selectively activated coating at the mapped location in apposition to identified cellular material of the tissue sample which is to be extracted.
  • the selectively activating coating is activated and impressed or impressed and activated to form an adhesive region on the transporting substrate for adhering to the identified cellular material.
  • identified cellular material adheres the transporting substrate at the mapped location.
  • coated tape 18 may have several differing configurations.
  • coated tape 18A is shown constructed from, e.g., polyester films such as Mylar ® having discrete and spaced apart coated spots 52 placed on coated side 19 of the tape. Additionally, sprocket holes 54 are shown for positioning the tape with precision in 13 combination with sprocket 56. Each different discrete and spaced apart coated spots 52 is separately activated at its coated center by the laser. Apposition to the tissue during or after laser action occurs by using a pressure plate.
  • a continuous strip including a transparent substrate is used to hold the film.
  • the continuous strip of transparent substrat'e or coated tape 18B has continuous central strip of activatable coating 58.
  • Adjacent bar codes 60 or other optical encoded identifiers could serve to identify the individual transferred samples, which samples can be either anywhere along the continuous central strip of activatable coating 58 or evenly spaced.
  • coated tape 18 transporting substrate with selectively coated spots or discrete and spaced apart coated spots 52 likewise can be used.
  • the substrate can be locally removed from the remainder of the substrate tape by punching or cutting off the local regions of activated coating.
  • punch 62 is shown separating desired sample sections 34 on punched film sections 64. This will leave attached identified cellular material or desired sample sections 34 localized to small pieces of transporting substrate on punched film sections 64. These small pieces of transporting substrate can be placed into a specific chamber or vessel such as test tube 66 for further processing and molecular analysis.
  • activatable coating 68 is attached to pull tabs 70 on coated tape 18C. As is apparent, by removal of pull tabs 70, sample gathered to activatable coating 68 is likewise removed.
  • coated tape 18D is shown being gathered to take up roll 72.
  • covering tape 74 covers over and helps in the preservation of the remaining sample on an interleave basis . Sample thus collected can be preserved. 14
  • FIG. 8 another embodiment of this invention is set forth.
  • Individual pieces of selectively activatable coating 76 are mounted at the ends of spoke arrays 78 attached to and projecting from central common support, such as central hub 80.
  • Spoke actuator 82 selectively depresses each spoke 79 of spoke array 78.
  • each spoke with its selectively activatable coating at the end acts as the transporting substrate and is sequentially indexed through the center of the microscope field from central hub 80.
  • the ends of the spokes serve as a pressure plates, contacting the tissue with the activatable coating at the ends of the spokes.
  • the selectively activatable coat can be actuated when the spoke is deflected, e.g., by a solenoid, (one spoke at a time, similar to a daisy wheel printer) . After activation, the force on the spoke is removed and the spoke indexed, allowing the spoke to lift off the slide with the identified cellular material adhered to the spoke. Thereafter, the spoke array indexes the next piece of film into place and the stage can be moved to the next tissue target.
  • a solenoid one spoke at a time, similar to a daisy wheel printer
  • central hub 80 is utilized.
  • a comb like structure, or a flexible backing can be used for the substrate support .
  • a preferred variant of this approach is to use comb shaped array 80 (i.e, target film spots are in a linear array attached to a linear carriage by parallel spokes perpendicular to the array axis rather than the radial spokes of a "daisy wheel") .
  • Minute spot of activatable coating 88 is placed a on the undersurface at the end of each comb finger 84.
  • Small solenoid 86 deflects the finger so its end, which acts as a pressure plate, contacts .the tissue in sample S in the center of the field.
  • the solenoid force is removed, the finger returns to its undeflected position (lifting off the slide the tissue adhered to the film) , and the comb is linearly indexed so a different finger/piece of film is in the center of the field and a different tissue target can be moved in place.
  • Optical marks 90 on the base of the comb could 15 identify the samples. This technique would allows extremely small and precise sample extraction.
  • spoke 79 with the attached identified cellular material can be broken off, and the broken off spoke portion 92 deposited into a container such as test tube 66 capsule for collection and further processing. Braking off can occur through breaker solenoid 94 at some convenient point along its length. It is possible with the disclosed invention and known technology to separate visualization and the requisite laser capture micro dissection.
  • slide 32 is shown having sample S viewed by microscope M. Slide 32 is mounted on stage 96 indicated by x-translation arrow 98 and y-translation arrow 99.
  • the typical sample S will include number sample locations as schematically indicated by x ⁇ y*** . through x 4 ,y 4 . These stage positions, when noted under observation, would be placed in conventional computer memory commonly provided with such stages .
  • Fig. 13C it will be seen that laser capture micro dissection is then remote.
  • the apparatus of Fig. 2 schematically shown by film strip laser capture micro-dissection apparatus A is shown in the process of sequential gathering of samples at x- ⁇ y- L through x 4 , y 4 (See Fig. 13B) .
  • the collection sample can extend to smaller samples.
  • utilizing techniques of laser capture microdissection it has been possible to collect samples down to a single cell in size (about five (5) microns reduced down to one (1) micron) or subcellular targets. With such small samples, correspondingly reduced analysis volume is required.
  • reagents for some analyses are extremely expensive; the smaller the sample and reagent used, the more economical the test. Laser capture 16 micro dissection is especially adaptable to such circumstances .
  • the laser activation allows targeting of elements on a complex tissue slide down to optical diffraction limits (ie. approximately 1 micron). Further, by precise positioning of film and substrate after each transfer, multiple small tissue elements can be precisely concentrated so that they comprise enough homogenous material to allow accurate subsequent processing (such as molecular analysis) .
  • concentration works best where contact between the sample and the tape does not occur before activation. Presuming that precise and accurate tracking of previously accumulated material is made, sample previously collected to the tape will not be physically impacted by sample currently being collected. At the same time, this subsequently collected sample will be adjacent the previously collected sample. By repeating this collection to adjacent portions of a substrate, concentration of the particular cells to be analyzed can occur, even though such cells were not naturally concentrated in situ.
  • selected cellular areas should not overlap. If overlapping occurs, sample can be lost. Further, with imprecise collection, the gap utilized could change effecting the predictability of the collection parameters. For example, the dosimetry of the laser radiation could be changed.
  • the spacing between the tape and sample can be in the broad range of 2 to 20 microns, in the intermediate range of 5 to 15 microns, and preferably about 10 microns when the thickness of the activatable coating is in the preferable range of 100 microns. Thicknesses of the activatable film in the range 17 from 20 to 200 microns can be used. As a practical limit, the thickness should be sufficient that the coating when activated has the desired adhesive effect. As a practical limit, it is presently believed that the coating should exceed 10 microns. Referring to Fig. 14A, slide 32 is shown with sample
  • Fig. 14B laser light source L is shown passing through slide 32, sample S, and onto coated side 19. As schematically shown by the broken lines of Fig. 14B, it will be understood that the laser light source L can be incident from an opposite direction.
  • Coated side 19 becomes locally activated and swells in dimension.
  • Activated region 112 extends downward at extended column 114 and at sample exposed portion 116, fastens to selected cellular material C.
  • extended column 114 shrinks, substantially back to the original profile of coated side 19. At this time, the contracting extended column 114 with selected cellular material C separates from sample S. (See Fig. 14C).
  • coated tape 18 can be transported so that a new segment of the coated tape overlies a targeted portion of a sample S on slide 32.
  • a new slide 32A can be introduced; thus several differing samples S from differing slides can be utilized for the collection and concentration of selected cellular material C.
  • tissue from two slides 32 and 32A ends up side-by-side on coated tape 18.

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  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Sampling And Sample Adjustment (AREA)
  • Microscoopes, Condenser (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

L'invention concerne un procédé et un appareil permettant d'extraire, au moyen d'une microdissection par piégeage laser, de la matière cellulaire identifiée, à partir de zones aléatoires d'un échantillon de tissu pour réunir cette matière sur des zones désignées d'un substrat de transport. Ce procédé et cet appareil permettent un traitement ultérieur approprié. Un substrat de transport comprend une zone localisée identifiée destinée à recevoir la matière cellulaire identifiée. Au moins un segment d'un revêtement sélectivement activé est placé sur le côté du substrat de transport, en apposition à l'échantillon de tissu sur la zone localisée. Le substrat de transport et l'échantillon sont déplacés l'un par rapport à l'autre de manière à placer le revêtement sélectivement activé sur la zone localisée, en apposition à la matière cellulaire identifiée de l'échantillon de tissu destinée à être extraite. Par la suite, le revêtement sélectivement activable est activé puis imprimé, ou imprimé puis activé, de manière à former sur le substrat de transport une région d'adhérence à la matière cellulaire identifiée. Lorsqu'on retire le substrat de transport de l'échantillon de tissu, la matière cellulaire identifiée adhère au substrat de transport sur la zone localisée.
PCT/US1999/001987 1998-02-03 1999-01-29 Systemes de manutention mecanique destines a la microdissection par piegeage laser WO1999039176A1 (fr)

Priority Applications (5)

Application Number Priority Date Filing Date Title
AU24850/99A AU749378B2 (en) 1998-02-03 1999-01-29 Mechanical handling systems for laser capture microdissection
EP99904451A EP1053461A1 (fr) 1998-02-03 1999-01-29 Systemes de manutention mecanique destines a la microdissection par piegeage laser
JP2000529587A JP4344474B2 (ja) 1998-02-03 1999-01-29 レーザ捕捉顕微分析のための機械的処理システム
US09/601,559 US6720191B1 (en) 1998-02-03 1999-01-29 Mechanical handling systems for laser capture microdissection
CA002319589A CA2319589C (fr) 1998-02-03 1999-01-29 Systemes de manutention mecanique destines a la microdissection par piegeage laser

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US7348098P 1998-02-03 1998-02-03
US60/073,480 1998-02-03

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WO1999039176A1 true WO1999039176A1 (fr) 1999-08-05

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EP (1) EP1053461A1 (fr)
JP (1) JP4344474B2 (fr)
AU (1) AU749378B2 (fr)
CA (1) CA2319589C (fr)
WO (1) WO1999039176A1 (fr)

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000034757A1 (fr) * 1998-12-10 2000-06-15 The Government Of The Unites States Of America Represented By The Secretary, Department Of Health And Human Services Conceptions de microdissection par piegeage laser sans contact
WO2001073397A1 (fr) * 2000-03-27 2001-10-04 P.A.L.M. Microlaser Technologies Ag Dispositif collecteur destine a recueillir des objets dissous dans un produit, en particulier par rayonnement laser
US6743601B1 (en) 1998-12-10 2004-06-01 The United States Of America As Represented By The Department Of Health And Human Services Non-contact laser capture microdissection
WO2004045768A1 (fr) * 2002-11-20 2004-06-03 P.A.L.M. Microlaser Technologies Ag Porte-echantillons pour un dispositif de reception recevant des objets biologiques et systeme de microscope fonctionnant avec un tel porte-echantillons
US6908766B2 (en) * 2002-10-18 2005-06-21 Industrial Technology Research Institute Device and method for capturing biological tissues
US7075640B2 (en) 1997-10-01 2006-07-11 Arcturus Bioscience, Inc. Consumable for laser capture microdissection
DE102005007793A1 (de) * 2005-02-14 2006-08-31 Laserinstitut Mittelsachsen E.V. Verfahren zur Herstellung von Körpern mit Laserstrahlen mindestens eines Lasers aus für die verwendete Laserwellenlänge transparenten Materialblöcken
DE102006051460A1 (de) * 2006-10-31 2008-05-08 P.A.L.M. Microlaser Technologies Gmbh Vorrichtung, Verfahren und Bandmaterial zum Aufsammeln und Transportieren von Probenmaterial
WO2008052836A1 (fr) * 2006-10-31 2008-05-08 Robert Bosch Gmbh Procédé de reconnaissance d'impuretés sur une surface
GB2550526A (en) * 2010-02-25 2017-11-22 Prairie Ventures Llc System and method for anatomical pathology sample handling, storage, and analysis

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2714803A1 (fr) * 2008-02-25 2009-09-03 Bizpac (Australia) Pty Ltd Appareil d'echantillonnage par perforation et procede associe
JP6611610B2 (ja) * 2013-10-07 2019-11-27 国立大学法人名古屋大学 レーザーマイクロダイセクション装置、該レーザーマイクロダイセクション装置を含む分析装置及びマイクロチップの製造方法
KR20200075814A (ko) * 2017-08-15 2020-06-26 옴니옴 인코포레이티드 화학적 및 생물학적 분석물의 검출에 유용한 스캐닝 장치 및 방법

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997013838A1 (fr) 1995-10-10 1997-04-17 The Government Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services Isolation d'un materiau sous visualisation microscopique
WO1999000658A1 (fr) * 1997-06-27 1999-01-07 The Government Of The United States Of America, Systeme de film adhesif a geometrie convexe pour microprelevement par laser

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997013838A1 (fr) 1995-10-10 1997-04-17 The Government Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services Isolation d'un materiau sous visualisation microscopique
WO1999000658A1 (fr) * 1997-06-27 1999-01-07 The Government Of The United States Of America, Systeme de film adhesif a geometrie convexe pour microprelevement par laser

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
BONNER R F ET AL: "LASER CAPTURE MICRODISSECTION: MOLECULAR ANALYSIS OF TISSUE", SCIENCE, vol. 5342, no. 278, 21 November 1997 (1997-11-21), pages 1481, 1483, XP002060626 *

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7221447B2 (en) 1997-10-01 2007-05-22 Molecular Devices Corporation Consumable for laser capture microdissection
US7075640B2 (en) 1997-10-01 2006-07-11 Arcturus Bioscience, Inc. Consumable for laser capture microdissection
AU760200B2 (en) * 1998-12-10 2003-05-08 Government Of The United States Of America, As Represented By The Secretary Of The Department Of Health And Human Services, The Designs for non-contact laser capture microdissection
US6743601B1 (en) 1998-12-10 2004-06-01 The United States Of America As Represented By The Department Of Health And Human Services Non-contact laser capture microdissection
WO2000034757A1 (fr) * 1998-12-10 2000-06-15 The Government Of The Unites States Of America Represented By The Secretary, Department Of Health And Human Services Conceptions de microdissection par piegeage laser sans contact
US7044008B1 (en) 2000-03-27 2006-05-16 P.A.L.M. Microlaser Technologies Ag Collecting device for collecting objects that are dissolved out of a mass, especially by means of laser radiation
WO2001073397A1 (fr) * 2000-03-27 2001-10-04 P.A.L.M. Microlaser Technologies Ag Dispositif collecteur destine a recueillir des objets dissous dans un produit, en particulier par rayonnement laser
DE10015157A1 (de) * 2000-03-27 2001-10-18 P A L M Gmbh Verfahren zur Bearbeitung einer biologischen Masse und Steuersystem für eine Vorrichtung zur Bearbeitung einer biologischen Masse
US6908766B2 (en) * 2002-10-18 2005-06-21 Industrial Technology Research Institute Device and method for capturing biological tissues
DE10254229A1 (de) * 2002-11-20 2004-06-17 P.A.L.M. Microlaser Technologies Ag Positioniervorrichtung zum Positionieren eines Positionierobjekts
DE10254229B4 (de) * 2002-11-20 2004-10-28 P.A.L.M. Microlaser Technologies Ag Positioniervorrichtung zum Positionieren einerAuffangvorrichtung eines Laser-Mikrodissektionssystems
WO2004045768A1 (fr) * 2002-11-20 2004-06-03 P.A.L.M. Microlaser Technologies Ag Porte-echantillons pour un dispositif de reception recevant des objets biologiques et systeme de microscope fonctionnant avec un tel porte-echantillons
US8431078B2 (en) 2002-11-20 2013-04-30 Carl Zeiss Microscopy Gmbh Sample holder for a reception device receiving biological objects and microscope system designed to operate using one such sample holder
DE102005007793A1 (de) * 2005-02-14 2006-08-31 Laserinstitut Mittelsachsen E.V. Verfahren zur Herstellung von Körpern mit Laserstrahlen mindestens eines Lasers aus für die verwendete Laserwellenlänge transparenten Materialblöcken
DE102005007793B4 (de) * 2005-02-14 2013-03-07 Lim Laserinstitut Mittelsachsen Gmbh Verfahren zum sowohl Herausarbeiten als auch Herauslösen freigeformter Körper mit Laserstrahlen mindestens eines Lasers aus für die verwendete Laserwellenlänge transparenten Materialblöcken
US8363213B2 (en) 2006-10-31 2013-01-29 Robert Bosch Gmbh Method for detecting impurities on a surface
WO2008052655A1 (fr) * 2006-10-31 2008-05-08 P.A.L.M. Microlaser Technologies Gmbh Dispositif, procédé et feuillard pour ramasser et transporter un matériau d'échantillon
WO2008052836A1 (fr) * 2006-10-31 2008-05-08 Robert Bosch Gmbh Procédé de reconnaissance d'impuretés sur une surface
DE102006051460A1 (de) * 2006-10-31 2008-05-08 P.A.L.M. Microlaser Technologies Gmbh Vorrichtung, Verfahren und Bandmaterial zum Aufsammeln und Transportieren von Probenmaterial
GB2550526A (en) * 2010-02-25 2017-11-22 Prairie Ventures Llc System and method for anatomical pathology sample handling, storage, and analysis
GB2490453B (en) * 2010-02-25 2018-01-17 Prairie Ventures Llc System and method for anatomical pathology sample handling, storage, and analysis
GB2550526B (en) * 2010-02-25 2018-02-07 Prairie Ventures Llc System and method for anatomical pathology sample handling, storage, and analysis
US9989449B2 (en) 2010-02-25 2018-06-05 Prairie Ventures, L.L.C. System and method for anatomical pathology sample handling, storage, and analysis
US11079307B2 (en) 2010-02-25 2021-08-03 Prairie Ventures, L.L.C. System and method for anatomical pathology sample handling, storage, and analysis

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CA2319589C (fr) 2008-11-18
CA2319589A1 (fr) 1999-08-05
JP4344474B2 (ja) 2009-10-14
AU2485099A (en) 1999-08-16
AU749378B2 (en) 2002-06-27
JP2002502025A (ja) 2002-01-22

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