WO2018228508A1 - Revêtement de protection en paraffine de lame de microscope - Google Patents

Revêtement de protection en paraffine de lame de microscope Download PDF

Info

Publication number
WO2018228508A1
WO2018228508A1 PCT/CN2018/091383 CN2018091383W WO2018228508A1 WO 2018228508 A1 WO2018228508 A1 WO 2018228508A1 CN 2018091383 W CN2018091383 W CN 2018091383W WO 2018228508 A1 WO2018228508 A1 WO 2018228508A1
Authority
WO
WIPO (PCT)
Prior art keywords
paraffin
microscope slide
stain
biomaterial
inorganic
Prior art date
Application number
PCT/CN2018/091383
Other languages
English (en)
Inventor
Frederick Knute Husher
Jee Jong Shum
Original Assignee
Sunstone Scientific Limited.
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 Sunstone Scientific Limited. filed Critical Sunstone Scientific Limited.
Priority to JP2020519171A priority Critical patent/JP7284160B2/ja
Priority to EP18817800.8A priority patent/EP3639078A4/fr
Priority to CN201880039119.6A priority patent/CN110741301B/zh
Priority to KR1020207001289A priority patent/KR102368836B1/ko
Publication of WO2018228508A1 publication Critical patent/WO2018228508A1/fr
Priority to US16/715,715 priority patent/US11662564B2/en
Priority to US18/302,495 priority patent/US20230280578A1/en

Links

Images

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B1/00—Optical elements characterised by the material of which they are made; Optical coatings for optical elements
    • G02B1/10—Optical coatings produced by application to, or surface treatment of, optical elements
    • G02B1/14—Protective coatings, e.g. hard coatings
    • 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 a shield coating on a microscope slide.
  • the present invention particularly relates to an application of a paraffin layer to shield biomaterials and inorganic chemical deposits from microbial attack and oxidation. More particularly, the present invention relates to the application of a paraffin layer on a microscope slide as an exposure shield over deposited biomaterial reactive targets.
  • the paraffin shield layer blocks the biomaterial and chemical targets from exposure, which may lead to degradation due to oxidation and provides resistance to fungal growth, while using the existing stain processing steps to remove the paraffin from the shield and co-resident tissue section.
  • Paraffin wax in general, is a white or colorless soft solid, derived from petroleum, coal or oil shale, which consists of a mixture of hydrocarbon molecules containing between twenty and forty carbon atoms. It is solid at room temperature and begins to melt above approximately 37 °C (99 °F) ; its boiling point is >370 °C (698 °F) .
  • Common applications for paraffin wax include lubrication, electrical insulation, and candles; dyed paraffin wax can be made into crayons. It is distinct from kerosene and other petroleum products that are sometimes called paraffin.
  • paraffin wax is used to impregnate tissue prior to sectioning thin samples of tissue. Water is removed from the tissue through ascending strengths of alcohol (75%to absolute) and the tissue is cleared in an organic solvent such as xylene or one of the aliphatic substitutes, such as Xylol. The tissue is then placed in paraffin wax for a number of hours and then set in a mold with wax to cool and solidify; sections are then cut on a microtome.
  • paraffin is inherently known as containing anti-fungal and antibacterial agents which prevent the oxidation of the antigen sites and air borne acid/base degradation of the exposed sites.
  • the paraffin shield coating changes the viable life of the biomaterial and chemical targets from 3-5 days to 1-2 years enabling useful product life for the end user.
  • Removal of the embedding paraffin is also routine practice in order to expose the tissue section to subsequent Immunohistochemical (IHC) or Hematoxylin and Eosin (H&E) staining. Utilizing the same or similar paraffin formulation to shield other deposited materials on the same microscope slide ensures that no additional slide processing must take place before beginning the IHC or H&E staining.
  • IHC Immunohistochemical
  • H&E Hematoxylin and Eosin
  • CN204790174 U
  • CN204790174 U
  • the sealed chamber enables close microscopic examination of a sample without risk of it being damaged by the microscope optics or other handling.
  • Such implementation does not support encapsulation of the biomaterial directly or IHC processing, specifically the deparaffinization of the tissue section.
  • a paraffin coating over biomaterial and inorganic targets.
  • the biomaterial and inorganic targets are deposited onto microscope slide wherein the paraffin coating is selectively applied to cover just past the deposits as a shield.
  • the resultant coated microscope slide is post heated to melt and/or blend paraffin particles into monolithic surface coating sealing both the deposits and the slide surface surrounding the deposits.
  • the paraffin coating may be applied by spray coating, screen printing, ink jet methods, pad printing, and roller transfer printing etc.
  • the paraffin coated increases the shelf life of the tissue sections by preventing the oxidation of the antigen sites and air borne acid and or base degradation of the exposed sites.
  • Tissue blocks and sections cut from the tissue block are infused with paraffin that completely replaces the water content of the tissue’s cellular structure.
  • the paraffin inherently does not support bacterial and fungal growth, which ensures long term stability of the embedded biomaterials.
  • Target proteins deposited onto a microscope slide, glass or plastic present a rich food source to bacteria or fungal antagonists. Additionally, the protein’s antigen sites are susceptible to oxidation that effectively neutralizes the ability to bind detection antibodies to the protein. Many of the subsequent reaction binding sites are hydroxyls, which can become damaged through reactions with airborne acids and bases. Typically, slides containing protein deposits are stored at temperatures below what supports microbial growth. However, such a constraint limits the effective utilization of deposits. Additionally, the protein deposited slides are packaged in vacuum sealed containers to prevent oxidation damage. Unprotected protein deposited slides have an open-air shelf life between 2 and 5 days depending upon ambient temperature and airborne contaminate levels.
  • Targets that react uniquely with the chemical structure of inorganic stains are also susceptible to oxidation and reaction with airborne materials.
  • stain reactant targets include those for Hematoxylin, Eosin, and most of the special stains group.
  • the targets for these stains are designed to react singularly with but one stain.
  • Biological targets do not, for the most part, provide for such singularity but chemical constructs can. Therefore, the present invention provides a solution by providing a method for selectively coating the biomaterials as well as targets to shield them from any kind of degradation on any kind of microscope slide.
  • the paraffin is blended with a solvent to change the material state from solid to a liquid at room temperatures.
  • the blend uses Paraplast X-tra or equivalent with Xylene or a Green Xylene replacement: an Aliphatic solvent, for example Xylol to reduce the viscosity and slow down solidification following deposition.
  • the solvent may be selected but not limited to toluene, paint thinner, turpentine, or a 50: 50 mix of acetone &kerosene.
  • the solid paraffin is melted at no more than 75°C above the paraffin melt temperature until liquid, then slowly add an Aliphatic solvent until the saturation point is observed (solids are formed) . Allow the mixture to cool to 45°C and slowly add more Aliphatic until it is completely clear.
  • biomaterials may include but are not limited to proteins, peptides, conjugated proteins, protein coated beads, peptide coated beads, or conjugated coated beads and the special stain reactive end groups that uniquely capture a special stain material which react with the applied antibody and secondary stain reagents.
  • the paraffin layer may be deposited onto the adhesive coated microscope slide which include but are not limited to: spray, inkjet deposit, transfer printing (such as pad printing) , screen printing, and vapor deposition.
  • the paraffin layer must be less than 5um thick to ensure that during the paraffin removal step of the staining process, all the paraffin (shield and tissue section embedding) can be dissolved and carried away.
  • the criterion of the paraffin is:
  • a thin layer preferably about no thicker than 5 microns
  • ⁇ Have a melting temperature of less than 60°C and preferably less than 56°C and dissolves with exposure to xylene or xylol (aliphatic replacement) solvents, and
  • the tissue block embedding paraffin materials may include but not limited to TissuePrep &TissuePrep 2 by Thermo Fisher, melting temp 56°C, Paraplast &Paraplast plus by Leica, melting temp 56°C, Paraplast X-tra by Leica, melting temp 50-54°C.
  • Paraplast X-tra specifically incorporates butylated hydroxytoluene, a phenolic antioxidant, to reduce oxidation degradation of protein, peptide, and inorganic targets.
  • each is a blend of purified paraffin, synthetic polymers, and other materials to establish the melt temperature, hardness, and viscosity.
  • Inherent to paraffin is non-support of microbial growth.
  • the special stains may include but are not limited to: Alcian Blue, Analine Blue –Orange G Solution, Azan Stain, Bielschowsky silver stain, Brow &Benn -Gramm Stain, Cresyl Violet, DAB, Fontana Masson, Gordon and Sweet's silver staining, Grocett's Methanamine silver method, Hall's Bilirubin stain, Jones Methanamine silver method, Luxol Fast Blue, Luxol Fast Blue --Cresyl Violet, Mucicarmine (Mayer's Method) , Muller-Mowry colloidal Iron, Orange G, Nuclear Fast Red, PAS with Diastase Digestion, Periodic Acid Schiff (PAS) , Phosphotungstic Acid, Haematoxylin, Picro Sirius Red, Toluidine Blue Acidified, Trichrome --Gomoris One-Step, Trichrome --Masson's, Victoria Blue, Von Kossa, Weigert's Resorcin Fuchsin,
  • the targets may be selected but not limited to pigment colored deposits such as black and white, but can include any pigment color.
  • the microscope slide on which the aforementioned paraffin coat may be applied may be selected from but not limited to glass, plastic or any polymer material.
  • the paraffin maybe purified and water free.
  • the resultant microscope slide may be post heated to melt and/or blend paraffin particles into a monolithic surface coating sealing both the deposits and the slide surface surrounding the deposits.
  • the resultant microscope slide is heated after the paraffin is deposited to force the solvent out of the paraffin, ensuring that it returns to a hardened state. This must be done from the paraffin side of the slide, preferably using infrared light. Melting the paraffin from the top down ensures that the solvent is able to rise up and evaporate from the paraffin without encumbrance. The result is illustrated in Figure 1, which shows how the melted paraffin ensures a good seal at the edges of its deposit.
  • Figure 1 This is a cross section view of the selectively applied paraffin shield layer over the biomaterial and chemical deposits.
  • the paraffin has been melted after being deposited to drive out the solvent needed to make the paraffin liquid and seal the edge to the slide and/or slide adhesive coating.
  • Spray over the surface with low airflow A low liquid to air mix is preferred.
  • the mixture is sprayed onto the slide, through a mask, to cover the PRS targets. Typically takes 1-2 passes to form a layer ⁇ 5 microns thick.
  • the paraffin mixture reservoir and spray head are both heated to slightly higher than 56°C to ensure the paraffin is a fluid and will remain as a fluid while in flight from the spray head to the slide. Spray coverage from the head is nominally 0.375” in width, but a mask may be used for smaller shield areas.
  • a post infrared reheat ensures 100%sealing.
  • the stainless steel screen will be heated by passing an electric current through the wires of the screen between two parallel sides.
  • the temperature of the screen needs to be slightly below the paraffin melt temperature so that paraffin does not weep through to the bottom side of the screen. Basically, the paraffin behaves more as a paste than a liquid.
  • the PRS will need a reheat cycle to ensure 100%sealing.
  • the inkjet head needs to have an integrated heater within the print head to keep the paraffin in a liquid state.
  • a post reheat cycle on the slide will ensure 100%sealing.
  • a heated roller pulls up a film of paraffin from a heated reservoir onto the roller.
  • the roller then transfers a film of paraffin onto the slide much in the same fashion as a painting a wall with a napped roller.
  • a post reheat cycle on the slide will ensure 100%sealing.

Landscapes

  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Optics & Photonics (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Urology & Nephrology (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Hematology (AREA)
  • Food Science & Technology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Biotechnology (AREA)
  • Medicinal Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Pathology (AREA)
  • Sampling And Sample Adjustment (AREA)
  • Microscoopes, Condenser (AREA)

Abstract

L'invention concerne un revêtement de protection disposé sur une lame de microscope. En particulier, l'invention concerne l'application sélective d'une couche de paraffine pour protéger des biomatériaux et des dépôts de produits chimiques inorganiques contre une attaque microbienne et une oxydation. Plus particulièrement, l'invention concerne également l'application d'une couche de paraffine sur une lame de microscope en tant que protection contre une exposition sur des cibles réactives de biomatériaux déposées. La couche de protection en paraffine bloque les cibles de biomatériaux et de produits chimiques contre une exposition, qui peut conduire à une dégradation due à l'oxydation et fournit une résistance à la croissance fongique, tout en faisant appel aux étapes de traitement de taches existantes pour éliminer la paraffine de la protection et de la section de tissu résidant au même emplacement.
PCT/CN2018/091383 2017-06-15 2018-06-15 Revêtement de protection en paraffine de lame de microscope WO2018228508A1 (fr)

Priority Applications (6)

Application Number Priority Date Filing Date Title
JP2020519171A JP7284160B2 (ja) 2017-06-15 2018-06-15 パラフィン保護コーティングを有する顕微鏡スライドの作成方法
EP18817800.8A EP3639078A4 (fr) 2017-06-15 2018-06-15 Revêtement de protection en paraffine de lame de microscope
CN201880039119.6A CN110741301B (zh) 2017-06-15 2018-06-15 用于显微镜载片的石蜡屏蔽涂覆层
KR1020207001289A KR102368836B1 (ko) 2017-06-15 2018-06-15 현미경 슬라이드용 파라핀 차폐 코팅
US16/715,715 US11662564B2 (en) 2017-06-15 2019-12-16 Paraffin shield coating for microscope slide
US18/302,495 US20230280578A1 (en) 2017-06-15 2023-04-18 Paraffin shield coating for microscope slide

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US201762520319P 2017-06-15 2017-06-15
US201762520169P 2017-06-15 2017-06-15
US62/520,319 2017-06-15
US62/520,169 2017-06-15

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2018/091685 Continuation WO2018228574A1 (fr) 2017-06-15 2018-06-15 Lame de microscope avec cibles noire et blanche intégrées

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US16/715,715 Continuation US11662564B2 (en) 2017-06-15 2019-12-16 Paraffin shield coating for microscope slide

Publications (1)

Publication Number Publication Date
WO2018228508A1 true WO2018228508A1 (fr) 2018-12-20

Family

ID=64660789

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2018/091383 WO2018228508A1 (fr) 2017-06-15 2018-06-15 Revêtement de protection en paraffine de lame de microscope

Country Status (5)

Country Link
EP (1) EP3639078A4 (fr)
JP (1) JP7284160B2 (fr)
KR (1) KR102368836B1 (fr)
CN (1) CN110741301B (fr)
WO (1) WO2018228508A1 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113063945A (zh) * 2021-04-27 2021-07-02 河南赛诺特生物技术有限公司 一种免疫组化联合弹性纤维双重染色试剂盒、染色方法及应用

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4832855A (en) * 1987-01-14 1989-05-23 Idemitsu Petrochemical Company Limited Immersion oil for microscopy
JP2000010017A (ja) * 1998-06-17 2000-01-14 Nec Corp 顕微鏡観察用カバーグラスおよび観察方法
CN102540444A (zh) * 2011-12-20 2012-07-04 肇庆理士电源技术有限公司 一种显微镜样品观察面压平装置及样品压平方法
WO2013008142A1 (fr) * 2011-07-13 2013-01-17 Koninklijke Philips Electronics N.V. Support de filtre à milieu à changement de phase
CN104086085A (zh) * 2014-06-30 2014-10-08 常州大学 一种载玻片表面刻蚀规则微井阵列的方法
CN204790174U (zh) * 2015-06-30 2015-11-18 天津市康婷生物工程有限公司 一种不需盖玻片可直接用于制片的载玻片
CN106596546A (zh) * 2016-11-11 2017-04-26 中国科学院广州地球化学研究所 一种在显微镜载玻片上安置和筛选矿物单颗粒样品的方法

Family Cites Families (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH04131735A (ja) * 1990-09-25 1992-05-06 Chiyoda Manufacturing Co Ltd 一槽式包埋装置の運転方法
EP0822403B1 (fr) * 1996-08-02 1998-07-29 Milestone S.r.l. Procédé pour le traitement d'échantillons organiques
JP3413343B2 (ja) * 1997-04-21 2003-06-03 科学技術振興事業団 試料ブロックの製造方法及びその装置
JP4304765B2 (ja) * 1999-06-02 2009-07-29 和光純薬工業株式会社 非脱灰硬組織の包埋方法及びキット
JP2004515752A (ja) * 2000-09-15 2004-05-27 バイオジェネックス ラボラトリーズ ｉｎｓｉｔｕハイブリダイゼーションの向上
JP3678655B2 (ja) * 2001-01-24 2005-08-03 理想科学工業株式会社 孔版原紙の補強方法および孔版原紙の製版装置
US8283134B2 (en) * 2005-12-27 2012-10-09 Kyoto University Cassette for fixing, embedding and slicing biological tissues and method of using the cassette
JP4840695B2 (ja) * 2006-11-17 2011-12-21 株式会社カケンジェネックス 組織薄切片パラフィンマスキング方法、及び装置

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4832855A (en) * 1987-01-14 1989-05-23 Idemitsu Petrochemical Company Limited Immersion oil for microscopy
JP2000010017A (ja) * 1998-06-17 2000-01-14 Nec Corp 顕微鏡観察用カバーグラスおよび観察方法
WO2013008142A1 (fr) * 2011-07-13 2013-01-17 Koninklijke Philips Electronics N.V. Support de filtre à milieu à changement de phase
CN102540444A (zh) * 2011-12-20 2012-07-04 肇庆理士电源技术有限公司 一种显微镜样品观察面压平装置及样品压平方法
CN104086085A (zh) * 2014-06-30 2014-10-08 常州大学 一种载玻片表面刻蚀规则微井阵列的方法
CN204790174U (zh) * 2015-06-30 2015-11-18 天津市康婷生物工程有限公司 一种不需盖玻片可直接用于制片的载玻片
CN106596546A (zh) * 2016-11-11 2017-04-26 中国科学院广州地球化学研究所 一种在显微镜载玻片上安置和筛选矿物单颗粒样品的方法

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
See also references of EP3639078A4 *
YU , X. ET AL.: "Detachment of methacrylate-embedded sections from microscope slides can be prevented by heating on hotplate", JOURNAL OF HISTOLOGY & HISTOPATHOLOGY, vol. 1, 31 December 2014 (2014-12-31), pages 10, XP055556410, ISSN: 2055-091X *

Also Published As

Publication number Publication date
EP3639078A4 (fr) 2021-03-31
JP2020523613A (ja) 2020-08-06
CN110741301B (zh) 2021-11-05
CN110741301A (zh) 2020-01-31
JP7284160B2 (ja) 2023-05-30
KR102368836B1 (ko) 2022-02-28
KR20200036849A (ko) 2020-04-07
EP3639078A1 (fr) 2020-04-22

Similar Documents

Publication Publication Date Title
CN110753869B (zh) 用于特异染色的过程记录玻片
US3619254A (en) Thermometric articles and methods for preparing same
IE53883B1 (en) Preservative and fixative preparations for biological systems
WO2018228508A1 (fr) Revêtement de protection en paraffine de lame de microscope
US20090170152A1 (en) Tissue Conditioning Protocols
US20200116989A1 (en) Microscope slide
EP3733277B1 (fr) Substrat à motifs présentant des zones hydrophiles et hydrophobes
CN112839911A (zh) 可烧除保护性涂层
JPH04325446A (ja) 撥水性酸化物皮膜およびその形成法
CN105938065B (zh) 操作速度快的组化笔在细胞爬片免疫组化检测中应用
Kohlmeyer et al. Permanent microscopic mounts
La Russa et al. A scientific approach to the characterisation of the painting technique of an author: the case of Raffaele Rinaldi
WO2020075137A1 (fr) Lame d'enregistrement du processus de coloration et procédé d'utilisation associé
KR102342993B1 (ko) 면역 조직 화학적 착색을 위한 공정 기록 슬라이드
DE2635438C3 (de) Verfahren zum gleichzeitigen Anfärben und Versiegeln einer auf einen mikroskopischen Objektträger aufgebrachten und luftgetrockneten biologischen Probe
WO2018181483A1 (fr) Lame de verre, et solution pour produire ladite lame de verre
DE102008011444A1 (de) Thermochrom beschichtete Substrate und Verfahren zu deren Herstellung
Arluison et al. Cellular localization of RNA degradation and processing components in Escherichia coli
JPH11310869A (ja) 薄膜形成材料および薄膜形成方法
WO1996023041A1 (fr) Pulverisation et revetement thermoplastiques, et procede
US6168657B1 (en) Adhesive varnish to receive powdered pigments
Fink A solvent-free coating-procedure for the improved preparation of cryostat sections in light microscope histochemistry
de Ghetaldi et al. In-depth examination and analysis of Domenico Cresti's oil on wall paintings in Santa Maria della pace in Rome
JP2000309749A (ja) 塗料組成物
Dittus et al. Residues from cyclododecane consolidation following desalination

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 18817800

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2020519171

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2018817800

Country of ref document: EP

Effective date: 20200115