WO2000008511A1 - Lames a zones de reaction definies par des barrieres hydrophobes - Google Patents

Lames a zones de reaction definies par des barrieres hydrophobes Download PDF

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Publication number
WO2000008511A1
WO2000008511A1 PCT/US1999/013382 US9913382W WO0008511A1 WO 2000008511 A1 WO2000008511 A1 WO 2000008511A1 US 9913382 W US9913382 W US 9913382W WO 0008511 A1 WO0008511 A1 WO 0008511A1
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WO
WIPO (PCT)
Prior art keywords
slide
reaction zone
recited
approximately
slides
Prior art date
Application number
PCT/US1999/013382
Other languages
English (en)
Inventor
Maria Fu
B. J. Kerns
Original Assignee
Biogenex Laboratories
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 Biogenex Laboratories filed Critical Biogenex Laboratories
Priority to EP99928640A priority Critical patent/EP1101143A1/fr
Priority to JP2000564087A priority patent/JP2002526785A/ja
Publication of WO2000008511A1 publication Critical patent/WO2000008511A1/fr

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Classifications

    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/34—Microscope slides, e.g. mounting specimens on microscope slides

Definitions

  • the present invention relates to microscope slides and, more particularly, to microscope slides with at least one reaction zone which is bounded by a hydrophobic border and which has an affinity for the samples to be analyzed.
  • Cell antigens have been recognized as important markers indicating the presence of pathological conditions in patients. Methods have been described in the literature for the detection of such antigen markers on cells, based on the visualization of the reaction of the antigens with the corresponding antibodies by fluorescence, gold particles or enzymatic development of visible dyes. Such methods are generally referred to as immunostaining techniques.
  • the reaction steps can be carried out on suspended cells, where evaluation takes place by means of transmission measuring devices, or by means of a microscope after transfer of the stained cells to a microscope slide.
  • Immunostaining techniques are also performed on tissue specimens take from a patient, for example by means of a biopsy. Such tissue specimens are routinely mounted on slides for ease of handling and analysis. Because of the manner in which slides with tissue specimens are prepared (see Elias, J., "Immunohistopathology: A Practical Approach to Diagnosis” ASCO Press, 1990, pp. 3-4, for examples of such preparation), the size and/or location of a tissue sample on a microscope slide can vary considerably within a relatively large area of the slide. In order to apply a stain to the correct location on a slide and to provide rinsing and other manipulation steps at appropriate times and in proper amounts, until recently all such staining operations were carried out by hand.
  • the literature contains descriptions of techniques for fixing the cells employed for such detection methods on the slide surface, for example by producing delimited reaction fields, which are in a position to fix the preparations, such as cells, to be examined.
  • it is known to cover the surface of a slide with a plastic film, in which one or more open regions have been created.
  • the slide surface exposed by these apertures in the plastic film is then coated with a substance, for example with poly-L-lysine (Transplantation 25(6):331-334 (1978)), which is able to adhere the cell preparation to the slide.
  • the present invention provides a slide for the immobilization and subsequent analytical processing of biological specimens.
  • the present invention provides a slide comprising a slide base defining a substantially rectangular planar upper surface and at least one reaction zone defined on the upper surface comprising a hydrophobic barrier circumscribing substantially the entire boundary of the reaction zone.
  • the barrier by virtue of being applied to the slide base will project upwardly from the upper surface of the slide to a height sufficient to prevent a predetermined amount of an aqueous solution deposited on the reaction zone from crossing the boundary of the zone.
  • the slide provides means for increasing the binding between the upper surface of the slide within the reaction zone and a biological specimen which is placed in contact with the upper surface. This binding will be sufficient to permit the specimen to be subjected to analytical processing steps as desired by the user.
  • the present invention is particularly adapted to the use of automated slide processing apparatus which utilize typically preprogrammed, repetitive motions and processing steps.
  • Figure 1 presents a top plan view of a slide in accordance with the present invention wherein the reaction zone consists of a single zone covering approximately
  • Figure 2 depicts a top plan view of a slide wherein the reaction zone covers approximately 50% of the slide.
  • Figure 3 depicts a top plan view of a slide in accordance with the invention which comprises a plurality of reaction zones provided by including two additional intermediate hydrophobic barriers in the slide of Figure 1 which subdivide the reaction zone into three separate zones of approximately equal dimensions.
  • the present invention provides a slide for the immobilization and subsequent analytical processing of biological specimens.
  • the present invention provides a slide comprising a slide base defining a substantially rectangular planar upper surface and at least one reaction zone defined on the upper surface comprising a hydrophobic barrier circumscribing substantially the entire boundary of the reaction zone.
  • the barrier by virtue of being applied to the slide base will project upwardly from the upper surface of the slide to a height sufficient to prevent a predetermined amount of an aqueous solution deposited on the reaction zone from crossing the boundary of the zone.
  • the slide provides means for increasing the binding between the upper surface of the slide within the reaction zone and a biological specimen which is placed in contact with the upper surface.
  • microscope slides are typically used in analytical processing of cell and tissue specimens and the dimensions of such slides have largely been standardized as approximately one inch wide by three inches long or, in metric units, 25 mm wide by 75 mm long by approximately 1 mm thick. Slides have been constructed from numerous different materials including glass and various plastics but glass slides have proven to be the most readily accepted for various properties including their strength, transparency, and resistence to scratching or other defects. Particularly preferred glass includes soda lime glass.
  • the present invention comprises in part a slide base defining a substantially rectangular planar upper surface.
  • substantially rectangular is meant that a series of slides of the present invention utilizing standardized dimensions will be perceived as rectangular and will very typically only within the confineds of the standard variations inherent in modern manufacturing techniques.
  • the upper and lower surface of a conventional slide will largely be interchangeable, once the various aspects of the present invention begin to manifest on the slide, the interchangeability of the upper and lower surface is lost and the upper surface of the slide is clearly defined.
  • the upper surface of the slide can be generally recognized as comprising two distinct regions: the reaction region and the labeling region.
  • the labeling region will be identified on the present slide as consisting of a region of up to approximately 25% of the planar upper surface of the slide and located at one end of the rectangle. Again, for purposes of ease in the use of automation, the labeling region can be viewed as an index region identifying a certain portion of the slide which is not directly involved in the processing of the biological specimen and the remaining portion of the slide will be standardized for recognition by the automated processing apparatus as a region potentially to be treated during the course of analytical processing.
  • the labeling region of the slide will typically consist of a labeling zone intended for identification of the biological specimen, its origin, the analytical processes to be conducted thereupon, and any additional information which the user considers important to identify in close conjunction with the specimen on the slide.
  • This labeling can be provided by a roughened surface treatment, termed "frosted glass" which enables the user to write information onto the slide as desired.
  • frosted glass which enables the user to write information onto the slide as desired.
  • such information can be provided by means of a label containing either verbal or machine readable information, such as, e.g., bar code encoded information.
  • the labeling region provides a means for unique identification of the slide and its contents.
  • the labeling region will also include sufficient area to identify, for example, the manufacturer of the slide or the processing apparatus whose operation the slide is intended to facilitate.
  • the present invention provides at least one reaction zone which will be defined on the upper surface of the slide.
  • the size of the reaction zones and their location on the slide surface In order to facilitate automated processing, and to conserve the expenditure of valuable and expensive analytical reagents, it is considered desirable to standardize the size of the reaction zones and their location on the slide surface.
  • the selection from this limited number of slides and the use of, for example, machine readable labels in the labeling zone will facilitate the operation of the automated processing apparatus and the aforementioned conservation in the use of valuable reagents.
  • the outer edges of at least a portion of the reaction zone will be substantially coterminous with the edges of the slide base providing the aforementioned upper surface of the slide.
  • a reaction zone is provided which covers approximately 2/3 of the upper surface of the slide leaving from 1/4 to 1/3 of the upper surface as definin the labeling region.
  • the reaction zone boundary will be substantially coterminous with the edges of the slide base on three sides and parallel with the fourth side of the slide base.
  • the reaction zone of this embodiment will be approximately 50 mm long and approximately 25 mm wide, assuming that the barrier member which defines the reaction zone will be substantially negligible in width.
  • the hydrophobic barrier member will substantially circumscribe the entire boundary of the reaction zone, desirably allowing only such gaps in continuity as are consistent with automated manufacturing processes. In the ideal case, no gaps will exist and the barrier member will be seen to completely circumscribe the boundary of the reaction zone.
  • the barrier member By virtue of its application to the slide base, the barrier member will have a certain height in that it will project upwardly from the upper surface of the slide base.
  • the magnitude of upward projection is considered to be relative insubstantial in that it is the properties of the barrier member itself rather any mechanical attributes which are intended to define the reaction zone and contain within its boundary any materials applied during the course of processing.
  • a further embodiment of the present invention is as depicted in Figure 2 in which the reaction zone comprises approximately 50% of the entire upper surface of the slide base. That is, again utilizing a standard slide with the aforementioned dimensions, the reaction zone in this embodiment will be approximately 30 mm long and approximately 25 mm wide. In this regard, a somewhat smaller region of the upper surface of the slide will be utilized for the immobilization of the biological specimen and any subsequent processing steps in automated analytical processing will require application of reagents and other solutions to a decreased area on the slide. In this manner, smaller biological specimens may be mounted in the more constrained region and less reagent coverage will be required in order to insure that the biological specimen has been treated.
  • FIG. 3 Yet another embodiment of the present invention is as depicted in Figure 3.
  • This embodiment comprises a plurality of reaction zones which are defined by the inclusion of two intermediate barrier members subdividing the reaction zone depicted in
  • Figure 1 into three separate zones of approximately equal dimension. That is, utilizing the standardized slide mentioned previously, each of the three separate zones will comprise a region approximately 15 mm long and approximately 25 mm wide. In this manner, a plurality of biological specimens can be mounted on the same slide and subject to the same, or different, analytical processing steps for purposes of direct comparison. Alternatively, it may simply be desirable to include three specimens of somewhat lesser size in order to increase the throughput of the automated processing apparatus. Although perhaps somewhat more cumbersome, it is contemplated that the information included in the labeling region, typically by resort to machine readable data, will be able to identify the three reaction zones separately by resort to the indexing function of the labeling region and in this regard, separate biological specimens and separate processing steps can be utilized and the information can be tracked appropriately.
  • the barrier member of the present invention which defines and circumscribes substantially the entire boundary of the reaction zone, will desirably have various properties which inhibit the migration of various materials applied to the reaction zone and substantially prevent cross contamination beyond the reaction zone boundaries.
  • One feature mentioned previously which could be expected to play a modest role in this barrier property would be the height of the barrier member projecting upwardly from the upper surface of the slide base. However, it is anticipated that this contribution would be relatively modest although difficult to gauge in any absolute sense.
  • the barrier member will comprise a hydrophobic material and the hydrophobic properties of the barrier member will be expected to perform the majority of the barrier function, particularly in view of the use of aqueous-based reagents and solutions in virtually all automated processing protocols.
  • hydrophobic and hydrophilic interactions thus enables the design of a reaction zone on the slide base which permits a predetermined quantity of reagent to be applied systematically throughout the reaction zone in anticipation that the reagent and/or aqueous solution will remain in a desired location and will not result in loss of reagent nor in cross-contamination between reaction zones on a single slide or between reaction zones on adjacent slides.
  • hydrophobic ink Although numerous hydrophobic compounds are well known in the art, it is presently considered desirable to make use of a hydrophobic ink to permit the reaction zone to be defined on the upper surface of the slide base by the use of various mechanical manufacturing techniques. Among such techniques include, for example, silk screen application or computerized control of a precise barrier member by utilization of for example, a computer controlled ink jet printer.
  • a desirable component of the hydrophobic ink will be a material such as Teflon ® a well known material having very substantial hydrophobic properties.
  • the ink will contain various non- volatile components, such as the Teflon ® and any pigments or other materials intended to promote uniformity and homogeneity in the applied barrier member.
  • the barrier member constructed in accordance with this preferred embodiment will, on the slide, consist of the non- volatile residue of the hydrophobic ink which is utilized in the creation of the barrier member.
  • S-902 gray a hydrophobic ink including Teflon ® available from Erie Scientific Company, Portsmouth, New Hampshire.
  • the various analytic protocols intended to be practiced upon the present slides typically call for at least one "washing" step, in which reagents or solutions previously deposited upon the reaction zone of the slide will be swept clear of the zone, often by resort to a washing solution or removal by other mechanical means.
  • a washing step using either a washing solution or other means, such as a blast of a gas such as air.
  • Yet another desirable feature of the present invention is the use of a means for increasing the binding between the upper surface of the slide, primarily within the reaction zone defined on the upper surface, and a biological specimen which is brought into contact with the upper surface and thus immobilized in the reaction zone for subsequent analytical processing.
  • a means for increasing the binding between the biological specimen and the upper surface of the slide within the reaction zone be sufficient to permit the specimen to be subjected to reasonable treatments commensurate with the accepted protocols for analytical processing.
  • the specimen should be prevented from detaching from the slide once it has been immobilized and should remain substantially permanently attached during subsequent processing, analysis, and storage.
  • the means for increasing the binding between the surface of the slide and the biological specimen can be included as a feature of the entire slide or can be restricted or limited to the reaction zone proper.
  • the selection of the appropriate technique for application will depend largely on considerations of efficiency of manufacture of the present slides and, possibly, the cost of the material used in providing the increased binding.
  • the region of the upper surface of the slide base within the reaction zone can be treated with an alkaline agent, such as with an alkali metal hydroxide or alkaline earth metal hydroxide, such as sodium hydroxide, or an alkaline detergent, which would render the treated surface of the slide base alkaline and thus provide a net positive charge.
  • an alkaline agent such as with an alkali metal hydroxide or alkaline earth metal hydroxide, such as sodium hydroxide, or an alkaline detergent, which would render the treated surface of the slide base alkaline and thus provide a net positive charge.
  • This net positive charge can have the effect of increasing the binding between the glass surface and the biological specimen or further agents can be employed to enhance or otherwise modify this binding effect.
  • Substances which have been shown to be suitable for this purpose are those which will adhere to the glass surface and bind to cells or proteins characteristic of the biological specimens both basic proteins and basic dyes such as poly-L-lysine, spermidine, polyarginine, and the like will bind to various components of biological cells or tissues, or isolated biological proteins as a result of the net negative charge on the cell or protein surface.
  • basic proteins and basic dyes such as poly-L-lysine, spermidine, polyarginine, and the like will bind to various components of biological cells or tissues, or isolated biological proteins as a result of the net negative charge on the cell or protein surface.
  • numerous substances are known which are capable of reacting with specific molecules including molecules located on the surface of cells and tissues, such as, for example, antibodies and including those which are specific to specified cell membrane antigens. This technique can be utilized for the isolation of specific proteins and cells where the slide of the invention is to be utilized not only for the immobilization of the biological specimen but for its identification and selection as well.
  • This technology is claimed to work well for adhering fresh cells, frozen tissue sections, and formalin fixed tissue specimens for various analytical processing protocols including immunoperoxidase, immunoalkalinephosphatase, color imetric and radiolabeled protocols for in situ DNA hybridization procedures. It has also been shown to be useful for use with frozen tissue sections, and other cytology preparations.
  • the specimens will bind to the slide and remain in place during staining procedures, microwave and chemical antigen retrieval procedures (subject of U.S. Patent Nos. 5,244,787 and 5,578,542 the entire contents of which are incorporated herein by this reference) as well as enzyme digestion, DNA denaturation and RNA hybridization procedures. It has however been noted that once the ADCELL-treated glass slides have been exposed to environmental agents, the net positive charge and thus the binding enhancement may dissipate over time.
  • Example 1 illustrates the preparation of slides in accordance with the present invention for the immobilization and subsequent analytical processing of biological specimens.
  • the slides will have at least one reaction zone defined by hydrophobic barrier circumscribing substantially the entire boundary of the zone.
  • Example 1 illustrates the preparation of slides in accordance with the present invention for the immobilization and subsequent analytical processing of biological specimens.
  • the slides will have at least one reaction zone defined by hydrophobic barrier circumscribing substantially the entire boundary of the zone.
  • a soda lime glass slide having the dimension 75 mm by 25 mm by 1 mm plus or minus 0.015 inches is provided.
  • the soda lime glass will consist of approximately 72.2% silicon dioxide, 14.3 % sodium oxide, 1.2% potassium oxide, 6.4% calcium oxide, 4.3% magnesium oxide, 1.2% aluminum oxide and trace amounts of ferric oxide and sulfur trioxide.
  • the slide will desirably be free of embedded foreign material such as bubbles, blisters and internal cloudiness and will also be free of rough or sharp cutting surfaces and edges.
  • one end of the slide as depicted in Figure 1 may be frosted to provide a labeling region for verbal or machine readable information.
  • a hydrophobic barrier region in the dimensions depicted in Figure 1 utilizing S902 gray hydrophobic ink containing Teflon ® (Erie Scientific Company, Portsmouth, NH).
  • the barrier member circumscribing the reaction zone will have a width of approximately 1 mm in the members defining the length of the reaction zone and approximately 2 mm in the members defining the width of the reaction zone.
  • the slide will have been treated with ADCELL adhesive cell treatment (Erie Scientific Company, Portsmouth, NH) to enhance its specimen binding characteristics.
  • ADCELL adhesive cell treatment (Erie Scientific Company, Portsmouth, NH) to enhance its specimen binding characteristics.
  • a soda lime glass slide having the dimension 75 mm by 25 mm by 1 mm plus or minus 0.015 inches is provided.
  • the soda lime glass will consist of approximately
  • the slide will desirably be free of embedded foreign material such as bubbles, blisters and internal cloudiness and will also be free of rough or sharp cutting surfaces and edges.
  • one end of the slide as depicted in Figure 2 may be frosted to provide a labeling region for verbal or machine readable information.
  • a hydrophobic barrier region in the dimensions depicted in Figure 2 utilizing S902 gray hydrophobic ink containing Teflon ® (Erie Scientific Company, Portsmouth, NH).
  • the barrier member circumscribing the reaction zone will have a width of approximately 1 mm in the members defining the length of the reaction zone and approximately 2 mm in the members defining the width of the reaction zone.
  • the slide will have been treated with ADCELL adhesive cell treatment (Erie Scientific Company, Portsmouth, NH) to enhance its specimen binding characteristics.
  • ADCELL adhesive cell treatment (Erie Scientific Company, Portsmouth, NH) to enhance its specimen binding characteristics.
  • a soda lime glass slide having the dimension 75 mm by 25 mm by 1 mm plus or minus 0.015 inches is provided.
  • the soda lime glass will consist of approximately 72.2% silicon dioxide, 14.3% sodium oxide, 1.2% potassium oxide, 6.4% calcium oxide, 4.3 % magnesium oxide, 1.2% aluminum oxide and trace amounts of ferric oxide and sulfur trioxide.
  • the slide will desirably be free of embedded foreign material such as bubbles, blisters and internal cloudiness and will also be free of rough or sharp cutting surfaces and edges.
  • one end of the slide as depicted in Figure 3 may be frosted to provide a labeling region for verbal or machine readable information.
  • a hydrophobic barrier region in the dimensions depicted in Figure 3 utilizing S902 gray hydrophobic ink containing Teflon ® (Erie Scientific Company, Portsmouth, NH).
  • the barrier member circumscribing the reaction zone will have a width of approximately 1 mm in the members defining the length of the reaction zone and approximately 2 mm in the members defining the width of the reaction zone.
  • the inclusion of two intermediate barrier members thus defining three substantially identical reaction zones is also depicted in Figure 3.
  • the slide will have been treated with ADCELL adhesive cell treatment (Erie Scientific Company, Portsmouth, NH) to enhance its specimen binding characteristics.
  • ADCELL adhesive cell treatment (Erie Scientific Company, Portsmouth, NH) to enhance its specimen binding characteristics.

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  • Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Optics & Photonics (AREA)
  • Sampling And Sample Adjustment (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Microscoopes, Condenser (AREA)

Abstract

Lames destinées à l'immobilisation et ensuite au traitement analytique d'échantillons biologiques présentant au moins une zone de réaction définie par une barrière hydrophobe entourant sensiblement toute la limite de ladite zone.
PCT/US1999/013382 1998-08-07 1999-06-14 Lames a zones de reaction definies par des barrieres hydrophobes WO2000008511A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP99928640A EP1101143A1 (fr) 1998-08-07 1999-06-14 Lames a zones de reaction definies par des barrieres hydrophobes
JP2000564087A JP2002526785A (ja) 1998-08-07 1999-06-14 疎水性防壁によって規定された反応区域を有するスライド

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US13066198A 1998-08-07 1998-08-07
US09/130,661 1998-08-07

Publications (1)

Publication Number Publication Date
WO2000008511A1 true WO2000008511A1 (fr) 2000-02-17

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EP (1) EP1101143A1 (fr)
JP (1) JP2002526785A (fr)
WO (1) WO2000008511A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010112016A1 (fr) 2009-03-30 2010-10-07 Dcs Innovative Diagnostik-Systeme Dr. Christian Sartori Gmbh & Co. Kg Procédé et dispositif de traitement de matière fixée à un support
WO2020092550A1 (fr) * 2018-11-01 2020-05-07 Leavitt Medical, Inc. Lames d'anatomopathologie ainsi que systèmes et méthodes associés

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1289904C (zh) * 2003-08-01 2006-12-13 博奥生物有限公司 一种微阵列反应装置及其应用
EA201491140A1 (ru) * 2011-12-09 2014-11-28 Дзе Скриппс Рисерч Инститьют Устройство, система и способ для идентификации циркулирующих опухолевых клеток
JP6362890B2 (ja) * 2014-03-18 2018-07-25 株式会社内田洋行 顕微鏡観察試料用具

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4705705A (en) * 1985-04-26 1987-11-10 Paul Marienfeld Kg Process for the production of slides with delimited reaction fields, and the slides produced by the process

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4705705A (en) * 1985-04-26 1987-11-10 Paul Marienfeld Kg Process for the production of slides with delimited reaction fields, and the slides produced by the process

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010112016A1 (fr) 2009-03-30 2010-10-07 Dcs Innovative Diagnostik-Systeme Dr. Christian Sartori Gmbh & Co. Kg Procédé et dispositif de traitement de matière fixée à un support
DE102009015596A1 (de) 2009-03-30 2010-10-21 Dcs Innovative Diagnostik-Systeme Dr. Christian Sartori Gmbh & Co. Kg Verfahren und Vorrichtung zur Behandlung von trägerfixiertem Material
WO2020092550A1 (fr) * 2018-11-01 2020-05-07 Leavitt Medical, Inc. Lames d'anatomopathologie ainsi que systèmes et méthodes associés

Also Published As

Publication number Publication date
JP2002526785A (ja) 2002-08-20
EP1101143A1 (fr) 2001-05-23

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