CN107614702A - The method for reducing evaporation at elevated temperatures - Google Patents
The method for reducing evaporation at elevated temperatures Download PDFInfo
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- CN107614702A CN107614702A CN201680023650.5A CN201680023650A CN107614702A CN 107614702 A CN107614702 A CN 107614702A CN 201680023650 A CN201680023650 A CN 201680023650A CN 107614702 A CN107614702 A CN 107614702A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6841—In situ hybridisation
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Abstract
A kind of method, including tissue or cell sample is contacted with reagent, the reagent includes having general formula X1‑R‑X2Evaporation reduce agent, wherein R is alkyl, alkenyl, alkynyl or the aromatic fractions of 1 or more carbon atom, can be substituted by oxygen, nitrogen or sulphur, and X1And X2By the independent part selected easily to be influenceed by hydrogen bond, and worked structure or cell sample.For handling the reagent of tissue or cell sample, wherein reagent includes evaporation reduction agent.
Description
The cross reference of related application
This application is originally non-provisional application, it is desirable in the Co-pending U.S. Patent Application submitted on April 15th, 2015
No.14/701, the priority of 043 more early submission date, and be incorporated herein by reference.
Technical field
Analytical reagent.
Background
Immunohistochemistry and determination techniques based in situ hybridization are widely used in medical diagnosis, such as diagnosis exception
Cell, such as those found in cancerous tumour, or another disease of diagnosis.Many measure, which are related into sample, adds heat
Measure, such as the sample in the reagent on slide.Determination step at elevated temperatures can cause to determine a large amount of of reagent
Evaporation.When using small size, denaturing samples, hybridization, washing and Aging Step and gained measure can be damaged by evaporation
Evil.
Detailed description of the invention
Disclosing a kind of aqueous solution includes the method that evaporation reduces the reagent of agent and uses the reagent, and its is operable
To reduce the evaporation of the reagent in tissue or cell sample processing.The representative processing of the reagent, which can wherein be used, to be included,
But it is not limited to, takes off paraffin, cell regulation/Antigen extraction (antigen retrieval)/cell senescence, peroxide/phosphoric acid
Enzyme blocks, probe denaturation, probe hybridization, washing, joint hybridization, antibody incubation, probe in detecting, chromogen precipitation and counterstain, its
The step of being related to elevated temperature (such as elevated temperature step in hybridization procedures in the original location).Solution can be used typically
In denaturation, washing or the hybridization step of nucleic acid (single or double chain probe) hybridization assays, or for being incubated, Antigen extraction or washing
Step, it uses antibody in immunohistochemistry or immunocytochemical stain.Suitable reagent can include other components,
Such as contribute in hybridization solution reagent to dissolve detergent/surfactant of diffusion.In one embodiment, available for group
Knit or cell processing reagent in evaporation reduce agent appropriate amount be under the conditions of industrial acceptable agent treatment will evaporation
Caused reagent loss is limited in 20% or less amount, and the agent treatment condition for example makes hybridization solution be subjected to 25 DEG C to 50
Rise temperature between DEG C, continue 10 minutes to 24 hours (for example, hybridization or incubation processes) or be subjected between 60 DEG C to 100 DEG C
Rise temperature, continue two to 90 minutes (for example, denaturation or Antigen extraction process).In one embodiment, evaporation subtracts
11% to 60% (by volume) of the appropriate amount scope of few agent in reagent composition (such as solution).
In one embodiment, reagent (composition) includes effectively reducing the evaporation reduction agent of evaporation capacity, and it includes logical
Formulas I：
X1-R-X2,
Wherein R is alkyl, alkenyl, alkynyl or the aromatic fractions of one or more carbon atoms (for example, in an embodiment
In one to six carbon atom, one to four carbon atom in another embodiment, if wherein should be understood that R is
The part of one carbon, then R is alkyl), it is by oxygen atom, sulphur atom and/or atom or nitrogen-atoms substitution (for example ,-OH ,-SH
Or-NH2Instead of the hydrogen atom for the one or more hydrocarbon for being defined as alkyl, alkenyl, alkynyl or aromatics) or can be by oxygen atom, sulphur
Atom and/or nitrogen-atoms interrupt (for example ,-C-O-C- ,-C-SH-C- ,-C-NH-C-).Moieties are containing for connecting X1
And X2Available tie point alkane.Alkenyl part is containing for connecting X1And X2Available tie point alkene.Alkynyl
Part is containing for connecting X1And X2Available tie point alkynes.Representational alkyl includes straight or branched alkyl (example
Such as, methyl, ethyl, propyl group, isopropyl), it can be further substituted or can not be further substituted.Representational alkenyl
Including straight or branched alkenyl (for example, vinyl, acrylic, isopropenyl).Representational alkynyl includes straight or branched
Alkynyl, it can be further substituted or cannot be further substituted (such as acetenyl, propinyl, isopropynyl).Aromatics
Part is the unsaturation ring of the stable atom of the interaction by forming the key of ring.The example of aromatic fractions is benzyl.Interrupt
Aromatic fractions example be pyridine (with nitrogen-atoms interrupt)；Furans (is interrupted) with oxygen atom；(used with thiophene in sulphur atom
Disconnected).
X in formula I1And X2Be independently selected as a part, one of those or two easily influenceed by hydrogen bonding and/or
Include electronegative atom.X1And X2Representative part include that the functional group of hydrogen bonding, such as, but not limited to hydroxylic moiety can be formed
(-OH)；Carbonyl moiety (- CO)；Amine moiety (- NH2)；Aldehyde part (- CHO)；(- Y, wherein Y are Cl to halogen moiety-, F-, Br-Or
I-)；Ether moiety (- OR1, wherein R1It is alkyl (for example, one arrives the alkyl of six carbon atom), it can be easy to be taken a group photo by hydrogen bond
Sound can form hydrogen bonding and/or the part substitution comprising electronegative atom)；Carboxy moiety (- COOR2, wherein R2It is hydrogen original
Son or alkyl (such as one to six carbon atom alkyl), it can be easy to that hydrogen bonding is influenceed or can formed by hydrogen bonding
And/or the part substitution comprising electronegative atom)；With amide moieties (- CONR3R4, wherein R3And R4Independently selected from hydrogen or alkane
Base (for example, one arrives the alkyl of six carbon atom), it can be easy to that hydrogen bonding and/or bag are influenceed or can formed by hydrogen bonding
Part substitution containing electronegative atom).For X1And X2The alkyl and ether, carboxyl and amide moieties are easy to by hydrogen bonding
Influence or can be formed the representative (R for substituting part of hydrogen bonding1, R2, R3And R4) include hydroxyl, carbonyl, amine, aldehyde, halogen, ether,
Carboxyl and amide moieties.
The example that suitable evaporation with formula I reduces agent includes ethylene glycol, glycine, serine, and isethionic acid is (different
Ethionic acid) (isoethionic acid), monoethanolamine, polyethylene glycol and 1,3-PD.In one embodiment, R is not
Substituted or interruption.Wherein R is unsubstituted or the representative of the evaporation reduction agent of interruption is ethylene glycol (HO (CH2)2OH), wherein R is
Ethyl, and X1And X2Be each hydroxylic moiety.Wherein X2It is further to be easy to that hydrogen bond is influenceed or can formed by hydrogen bonding
It is H that the evaporation of the partially substituted primary amide closed, which reduces agent example,2N-(CH2)3-NH(CH2)2COOH, wherein R are-(CH2)3-；X1
For-NH2And X2For-NH (CH2)2COOH.In one embodiment, suitable reagent includes the one or more with formula I
Evaporation reduces agent (for example, evaporation of the reagent including two or three of structural formula with formula I reduces the combination of agent).
It can be in a molecule free form (molecule) or its acceptable salt (compound) or ion that evaporation, which reduces agent,
Liquid.The example of salt is the halogen acid salt of molecule, such as hydrochloride.
In one embodiment, evaporation reduces agent and reduced as solute when it is mixed with one or more other reagents
The vapour pressure of solution.Such solute is probably molecule, and ion salt, ionic liquid, nonionics, it can show complete with water
Complete or partial solubility or compatibility and it can be reduced it is mixed into or the aqueous solution that otherwise introduces
Partial pressure.In addition, evaporation reduce agent can with or cannot with water formed azeotropic mixture.If evaporation reduces agent and forms azeotropic with water
Mixture, in one embodiment, solute form decline property azeotropic mixture mixture (negative effect azeotropic mixture) with water.
In general, when solvent and solute are mixed in this context, the gained chemical potential of solvent is lowered so that
The partial pressure of solvent molecule reduces, and relative to the solution of only solvent, has the tendency for being converted to gas phase reduced.Typically, it is aqueous
The partial pressure of solution (wherein solute is that non-volatile evaporation reduces agent) reclaimed water will be reduced, and this will cause the boiling point liter of mixture
Height, and the reduction in surface evaporation.
Hydrogen bond can occur when hydrogen atom in the molecule is connected to electronegative atom (such as oxygen, nitrogen or fluorine atom)
Close, and the close atom with positive electricity of molecule.The property of this electrostatic attraction has the intensity of the 5-30kJ/mol orders of magnitude, its
It is more stronger than Van der Waals force but weaker than covalent or ionic bonding.
In one embodiment, evaporation reduces agent as solute by being mixed below with water to reduce the vapour pressure of water：
Solution is created, the partial pressure of water therein reduces so that measure (for example, immuning tissue's determination method) than partial pressure if can not reduce
Situation be more effectively carried out.In one embodiment, solute is chosen to not leave residue or left minimum
Residue, the residue can disturb that well known to a person skilled in the art measure.
In one embodiment, the reagent that can be used to the cell or tissue sample of processing passes through combination foundation reagent
(base reagent) and evaporation reduce agent to prepare.Supplementary element can also be combined in suitable composition.The reagent can be by
It is added to aqueous buffer solutions to reduce the evaporation during elevated temperature.In one embodiment, the evaporation being combined subtracts
The amount of few agent limits any loss of the combination caused by evaporating to 20% or less, in another embodiment,
The amount that the evaporation being combined reduces agent limits any loss of the combination caused by evaporating to 10% or less.It is representative
Ground, evaporation reduce agent or evaporation reduce agent combination in the composition existing amount be 11% to 60% (by volume) amount,
Or it is compound containing non-ethylene glycol or the combination for reducing agent to reduce 1% to 60% amount of agent.In combination foundation reagent, (alkali tries
Agent), after evaporation reduces agent and any other component, composition can be mixed.
In one embodiment, be disclosed, in the measure of elevated temperature without paraffin or de- stone
Manipulable method in the processing of the tissue or cell sample of wax.In one embodiment, method is exercisable, to subtract
Few any evaporation based on aqueous reagent treatment.This method includes making tissue or cell sample contact include with formula I
The evaporation of formula reduces the reagent of agent (for example, as solute)：
X1-R-X2。
Such contact includes the sample being applied to reagent on slide.After sample is contacted with reagent, Ke Yigen
Sample is handled according to techniques known in the art.
In one embodiment, disclose, in the measure of elevated temperature without paraffin or de- paraffin
Tissue or cell sample processing in manipulable method.In one embodiment, method is exercisable, to pass through
Solute is added into solvent to reduce any evaporation based on aqueous treatment agent.The gained chemical potential of solvent reduces so that solvent
Molecule partial pressure reduce, and they become gas phase tendency it is smaller.Water in the aqueous solution that solute is nonvolatile matter
Partial pressure reduce, this will cause the elevation of boiling point and surface evaporation to reduce.
Embodiment
Embodiment 1：The Volume Loss caused by evaporation during oligonucleotide hybridization under different hybridization temperatures
The each and described slide 400 μ l hybridization solution (reagent) being applied in six specimen slides individually adds
Heat continues 30 minutes to different temperature.It was observed that steamed during hybridization step on the slide with hybridization solution
Hair.
Influence of the temperature of table 1. to evaporation
Embodiment 2：Evaporation during reducing oligonucleotide hybridization by different reagents.
The hybridization solution (reagent) that 400 microlitres there are different evaporations to reduce agent is applied in each specimen slide and added
Hot to 43 DEG C continue 30 minutes.One specimen slide includes reducing the hybridization solution of agent without evaporation.It was observed that 43
In DEG C hybridization step, notable more evaporations of hybridization solution occur to carry in the sample for reducing agent without evaporation containing hybridization solution
Slide, but agent is reduced to reduce by adding to evaporate.
The ability that reduction of the reagent that table 2. adds to hybridization solution (reagent) is evaporated
| Reagent | Average external volume loses (μ l) |
| Do not add | 55.0 |
| 20% ethylene glycol | 20.0 |
| 20% propane diols | 45.0 |
| 20% glycerine | 42.5 |
| 20% polyethylene glycol | 15.0 |
Embodiment 3：During hybridization evaporation is reduced using the ethylene glycol of various concentrations.
The hybridization solution (reagent) that 400 μ l are had to the evaporation reduction agent of the ethylene glycol of various concentrations is applied to each sample
Slide is simultaneously heated to 43 DEG C and continues 30 minutes.As a comparison, hybridization solutions of the 400 μ l without ethylene glycol is also applied to sample
Slide is simultaneously heated to 43 DEG C and continues 30 minutes.It was observed that reduce evaporation by adding ethylene glycol into hybridization solution.
It was found that the measure performance of the hybridization solution containing ethylene glycol is similar to the hybridization solution without ethylene glycol.
The Volume Loss of ethylene glycol (EG) is reduced (in terms of %) in the hybridization solution of table 3.
Embodiment 4：The repeatability of evaporated volume loss is reduced during hybridization.
There is the evaporation of certain concentration ethylene glycol to reduce the hybridization solution of agent 400 μ l and be applied to each specimen slide simultaneously
It is heated to 43 DEG C and continues 30 minutes.In order to compare, 400 μ l hybridization solutions are applied in each specimen slide and are heated to 43
DEG C continue 30 minutes.It was observed that reduce evaporation by adding ethylene glycol into hybridization solution.
Table 4. evaporates reduction caused by adding ethylene glycol
Embodiment 5：Evaporate and reduce during denaturation.
There is the evaporation of 20% ethylene glycol to reduce the hybridization solution of agent about 400 μ l to be applied in each specimen slide,
And it is heated to 92-98 DEG C and continues 2 minutes.In order to compare, hybridization solutions of the about 400 μ l without ethylene glycol is applied to each sample
On slide, and it is heated to 92-98 DEG C and continues 2 minutes.It was observed that reduce evaporation by adding ethylene glycol into hybridization solution.
Table 5. reduces evaporation in degenerative process by adding ethylene glycol into hybridization solution.
Embodiment 6：The reduction evaporated during antibody incubation step.
There is the evaporation of 20% ethylene glycol to reduce the primary antibody diluent (reagent) of agent about 400 μ l and be applied to each sample load
On slide, and it is incubated 30 minutes at a certain temperature.In order to compare, the primary antibody diluent that about 400 μ l do not have ethylene glycol is applied
On to each specimen slide and it is incubated 30 minutes at a certain temperature.It was observed that by adding ethylene glycol into antibody diluent
Reduce evaporation.
Table 6. reduces evaporation by adding ethylene glycol into antibody diluent
Embodiment 7：Evaporation during reducing oligonucleotide hybridization by different reagents.
The 400 μ l hybridization solutions that there are different evaporations to reduce agent are applied in each specimen slide and are heated to 43 DEG C
Continue 30 minutes.The hybridization solution that agent is reduced without evaporation is also put on into specimen slide.It was observed that betide 43
There is the slide of hybridization solution during DEG C hybridization step, but reduced by adding particular agent.
The evaporation that table 7. adds reduces the ability that reduction of the agent to dirt solution is evaporated
Embodiment 8：During hybridizing at different temperatures evaporation is reduced using ethylene glycol.
By 400 μ l have ethylene glycol hybridization solution (reagent) apply to each specimen slide and be heated to 35,37 or
40 DEG C continue 120 minutes, to compare, hybridization solutions of the 400 μ l without ethylene glycol are also put on into specimen slide, and heat
Continue 120 minutes to 40 DEG C.It was observed that reduce evaporation by adding ethylene glycol into hybridization solution.It was found that contain ethylene glycol
Measure performance of the hybridization solution in different hybridization temperatures is similar to the hybridization solution without ethylene glycol.
Reagent
Hybridization solution
| Chemicals | Ultimate density |
| SSC | 2x |
| Tris-HCl | 10mM |
| Dextran sulfate | 10% |
| Denhardt solution (Denhardt ' s solution) | 2x |
| Tween | 0.05% |
| Triton | 0.10% |
| Salmon sperm dna | 50μg/ml |
| Evaporation reduces agent | Change |
| Water | Change |
Primary antibody diluent
| Chemicals | Ultimate density |
| Sodium azide | 0.05% |
| Sodium chloride | 300mM |
| Disodium hydrogen phosphate | 8mM |
| Potassium dihydrogen phosphate | 2mM |
| Green colouring material (Green Color Dye) | 0.025% |
| BSA powder | 1% |
| Tris alkali | 10mM |
| Tris HCl | 40mM |
| Tween | 0.05% |
| Triton | 0.1% |
| Evaporation reduces agent | Change |
| Water | Change |
In superincumbent description, for purposes of explanation, many details have been elaborated to provide to embodiment party
The thorough understanding of case.It is clear, however, that for those skilled in the art, can be in some in these no details
In the case of put into practice other one or more schemes.It is not the limitation present invention to provide described specified scheme, but in order to say
The bright present invention.The scope of the present invention is not determined by specific examples provided above, and is only limited by claim afterwards.
In other cases, it is known that structure, equipment and operation have been shown as block diagram format or no details, to avoid fuzzy pair
The understanding of description.In appropriate circumstances, the end section of attached reference numeral or reference numeral is repeated to show in figure
Go out corresponding or similar element, it can optionally have similar characteristic.
It is also understood that in this manual to " embodiment ", " embodiment ", " one or more embodiment party
The reference of case " or " different embodiments ", such as mean that special characteristic can be included in the practice of the invention.Similarly,
It should be appreciated that in the de-scription, in order to simplify the disclosure and help to understand various inventive aspects, various features are together divided sometimes
Group is in single embodiment, figure or its description.However, this disclosed method is not necessarily to be construed as reflecting that invention requires every
The intention of the more features clearly described in individual claim.On the contrary, as the following claims reflect, creativeness side
Face may is that all features less than single disclosed embodiment.Therefore, claim is hereby bright afterwards in detailed description
Really it is incorporated in this detailed description, wherein independent embodiment of each claim independently as the present invention.
Claims (20)
1. a kind of method, including：
At least one evaporation with below general formula of tissue or cell sample contact is set to reduce agent：
X1-R-X2,
Wherein R is the moieties that can be substituted or be interrupted by oxygen, sulphur or nitrogen-atoms, alkenyl part, alkynyl moiety or aromatic fractions,
And X1And X2Independently be selected as comprising electronegative atom or the part that is easily influenceed by hydrogen bonding, and wherein when it is described at least
At least one reagent is with 1% or more when a kind of reagent is the combination of the non-reagent containing glycol or more than one reagent
Amount exist, or when at least one reagent is the reagent containing glycol at least one reagent with 11% by volume
Or more amount exist；With
Handle tissue or cell sample.
2. the method for claim 1 wherein the evaporation reduces amount presence of the agent with 60 volume % up to by volume.
3. the method for claim 1 wherein the processing includes elevated temperature.
4. the method for claim 1 wherein the processing includes hybridization.
5. the method for claim 1 wherein the processing includes going to hybridize.
6. the method for claim 1 wherein the processing includes washing step.
7. the method for claim 1 wherein the processing includes antigen binding.
8. the method for claim 1 wherein the processing includes Antigen extraction, cell regulation or cell senescence.
9. the method for claim 1 wherein X1And X2Independently it is selected so that X1And X2It is at least one containing selected from by following
The one or more of the group of composition：Hydroxylic moiety, carbonyl moiety, amine moiety, aldehyde part, halogen moiety, ether moiety, carboxy moiety
With the group of acid amides.
10. the method for claim 9, wherein X1And X2Independently it is selected as including at least one selected from the group consisted of
Kind part：- OH ,-CHO ,-OR1,-NH2,-COOR2With-CONH2, wherein R1It is alkyl and R2For hydrogen atom or alkyl.
11. the method for claim 1 wherein R is the alkyl that includes two or more carbon atoms, one of the carbon atom or
It is multiple to be substituted or can not be substituted by O, N or S by O, N or S.
12. the method for claim 1 wherein the reagent includes ethylene glycol.
13. the method for claim 1 wherein the reagent includes glycine, serine, isethionic acid (different ethionic acid)
(isoethionic acid), monoethanolamine, glycerine, polyethylene glycol, the combination of 1,3-PD or reagent.
14. the method for claim 1 wherein the reagent includes detergent or surfactant.
15. the method for claim 1 wherein the reagent includes the combination of evaporation reduction agent.
16. can be used to handle the reagent of tissue or cell sample, the reagent includes evaporation and reduces agent, when described at least one
The evaporation is reduced agent and deposited with 1% or more amount when kind of reagent is the combination of the non-reagent containing glycol or more than one reagent
, or the evaporation reduces agent with 11% or more by volume when at least one reagent is the reagent containing glycol
Amount exist, it is described evaporation reduce agent there is below general formula：
X1-R-X2
Wherein R is the moieties that can be substituted or be interrupted by oxygen, sulphur or nitrogen-atoms, alkenyl part, or alkynyl moiety, and X1And X2
Independently it is selected as comprising electronegative atom or the part easily influenceed by hydrogen bonding.
17. the reagent of claim 16, exist wherein the evaporation reduces agent with 60% amount up to by volume.
18. the reagent of claim 16, wherein X is selected independently1And X2So that X1And X2It is at least one containing selected from by following
The one or more of the group of composition：Hydroxylic moiety, carbonyl moiety, amine moiety, aldehyde part, halogen moiety, ether moiety, carboxy moiety
And amide moieties.
19. the reagent of claim 16, it includes detergent or surfactant.
20. the reagent of claim 16, it includes the combination that evaporation reduces agent.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/US2016/027945 WO2017180160A1 (en) | 2016-04-15 | 2016-04-15 | Methods to reduce evaporation during elevated temperature |
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| CN107614702A true CN107614702A (en) | 2018-01-19 |
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| JP (1) | JP6550471B2 (en) |
| CN (1) | CN107614702A (en) |
| AU (1) | AU2016402370B2 (en) |
| CA (1) | CA2983930C (en) |
| WO (1) | WO2017180160A1 (en) |
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| US20070048770A1 (en) * | 2005-07-15 | 2007-03-01 | Jaekel Robert W | Reagents and methods for processing biological samples |
| WO2012055981A1 (en) * | 2010-10-27 | 2012-05-03 | Glaxosmithkline Biologicals S.A. | Immunogenic compositions and methods for treating neurologic disorders |
| US20140242595A1 (en) * | 2013-02-22 | 2014-08-28 | Cellular Dynamics International, Inc. | Hepatocyte production via forward programming by combined genetic and chemical engineering |
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| WO1994002644A1 (en) * | 1992-07-17 | 1994-02-03 | Aprogenex, Inc. | In situ detection of nucleic acids using 3sr amplification |
| BR9306867A (en) * | 1992-07-17 | 1998-12-08 | Aprogenex Inc | Enrichment and identification of fetal maternal blood cells for in situ hybridization |
| US5500339A (en) * | 1993-03-30 | 1996-03-19 | Amersham Life Science, Inc. | DNA denaturation method |
| US6127188A (en) * | 1995-10-06 | 2000-10-03 | Mj Research, Inc. | Method and apparatus for controlling evaporation in histological procedures |
| AU766614B2 (en) * | 1998-09-03 | 2003-10-23 | Ventana Medical Systems, Inc. | Removal of embedding media from biological samples and cell conditioning on automated staining instruments |
| AUPQ257199A0 (en) * | 1999-08-31 | 1999-09-23 | Commonwealth Scientific And Industrial Research Organisation | Vaccine antigens of moraxella |
| US20040235104A1 (en) * | 2003-05-08 | 2004-11-25 | Millennium Pharmaceuticals, Inc. | Human SEF molecule and uses therefor |
| JP6214137B2 (en) * | 2012-07-17 | 2017-10-18 | 日本ハム株式会社 | Nucleic acid hybridization solution |
| EP3827820A1 (en) * | 2013-03-15 | 2021-06-02 | The Johns Hopkins University | Brivaracetam for improving cognitive function |
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- 2016-04-15 CN CN201680023650.5A patent/CN107614702A/en active Pending
- 2016-04-15 AU AU2016402370A patent/AU2016402370B2/en active Active
- 2016-04-15 WO PCT/US2016/027945 patent/WO2017180160A1/en active Application Filing
- 2016-04-15 JP JP2017554865A patent/JP6550471B2/en active Active
- 2016-04-15 CA CA2983930A patent/CA2983930C/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070048770A1 (en) * | 2005-07-15 | 2007-03-01 | Jaekel Robert W | Reagents and methods for processing biological samples |
| WO2012055981A1 (en) * | 2010-10-27 | 2012-05-03 | Glaxosmithkline Biologicals S.A. | Immunogenic compositions and methods for treating neurologic disorders |
| US20140242595A1 (en) * | 2013-02-22 | 2014-08-28 | Cellular Dynamics International, Inc. | Hepatocyte production via forward programming by combined genetic and chemical engineering |
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| CA2983930A1 (en) | 2017-10-19 |
| AU2016402370B2 (en) | 2023-07-06 |
| WO2017180160A1 (en) | 2017-10-19 |
| CA2983930C (en) | 2023-05-02 |
| JP6550471B2 (en) | 2019-07-24 |
| JP2018518941A (en) | 2018-07-19 |
| AU2016402370A1 (en) | 2018-01-18 |
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