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.

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• 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)
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• Microscoopes, Condenser (AREA)

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• 2018
  • 2018-06-15 JP JP2020519171A patent/JP7284160B2/ja active Active
  • 2018-06-15 WO PCT/CN2018/091383 patent/WO2018228508A1/en unknown
  • 2018-06-15 KR KR1020207001289A patent/KR102368836B1/ko active IP Right Grant
  • 2018-06-15 CN CN201880039119.6A patent/CN110741301B/zh active Active
  • 2018-06-15 EP EP18817800.8A patent/EP3639078A4/de active Pending

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