CN105823665A - Biological tissue sample preparation sleeve solution - Google Patents
Biological tissue sample preparation sleeve solution Download PDFInfo
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- CN105823665A CN105823665A CN201610319967.5A CN201610319967A CN105823665A CN 105823665 A CN105823665 A CN 105823665A CN 201610319967 A CN201610319967 A CN 201610319967A CN 105823665 A CN105823665 A CN 105823665A
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- sample preparation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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Abstract
The invention discloses a biological tissue sample preparation sleeve solution which comprises a fixed solution and a transparent solution. The fixed solution is prepared from, by volume, 45-50 parts of absolute ethyl alcohol, 20-25 parts of absolute methanol, 15 parts of polyethylene glycol 400, 10-20 parts of purified water and 0.03-0.05 part of a surface active agent; the transparent solution is prepared from, by volume, 90 parts of D-limonene, 10 parts of isopropyl alcohol and 0.03-0.05 part of a surface active agent. The biological tissue sample preparation sleeve solution can meet the requirement of environment-friendly pathological and histological reagents for tissue sample treatment in various hospital pathology departments and scientific research institutions, can meet the technical requirement for general pathological diagnosis, immunohistochemistry diagnosis and molecular biological diagnosis, and the tissue treatment effect is better than that of a traditional technology.
Description
Technical field
The present invention relates to the external diagnosis reagent in medical apparatus and instruments, particularly to a kind of biological organization sample preparation set liquid.
Background technology
The histopathologic birth history of existing more than 150 year, in all of hospital, as long as there being operation, by Hospital Pathological Department
Operation tissue samples is made histopathologic diagnosis by section.Routine paraffin wax embedding HE stained preparation is still current in order to process group
Knitting the most basic technology of sample, it is the basis of pathological diagnosis method, is also many new methods, the evaluation of new technique quality
Reference standard.At present, the whole nation or even the whole world are still using traditional method, it may be assumed that tissue samples is placed treatment fluid
In cylinder, pouring into successively or call in different disposal liquid, sample soaks different time by regulation successively in each reagent to be completed to process
Section, dyeing, mounting etc., omnidistance needs 30 several steps.Whole process use in a large number formaldehyde, dimethylbenzene, mercury oxide,
The toxic chemical reagent of ammonia, pathology technicians places oneself in the midst of toxic environment for a long time；Reagent tolerance is poor, operation is complicated, no
Easy to control, pathology slice-making quality is poor, impact diagnosis；Reagent consumption is big, too much discharges, serious environment pollution.The biggest
Number hospital reagent preparation voluntarily, lacks unified standard.Therefore the standardization of products used by pathology industry, standardization, low poison,
Low emission, is the inexorable trend of industry development.
Summary of the invention
It is an object of the invention to solve at least the above and/or defect, and the advantage that at least will be described later is provided.
It is a still further object of the present invention to provide a kind of biological organization sample preparation set liquid, can meet situation of all-level hospitals Pathology Deparment and
The environment-friendly type pathological tissue reagent that tissue samples is processed by scientific research institutions, can meet routine pathology diagnosis, immuning tissue
Learning diagnostic techniques, diagnostic technique in molecular biology requirement, the effect processing tissue is better than conventional art.
To this end, the technical scheme that the present invention provides is:
A kind of biological organization sample preparation set liquid, including fixative and transparent liquid, in terms of volume parts,
Described fixative includes dehydrated alcohol 45～50 parts, absolute methanol 20～25 parts, PEG400 15 parts, pure
Change water 10～20 parts and surfactant 0.03～0.05 part；
Described transparent liquid includes D-hesperidene 90 parts, isopropanol 10 parts and surfactant 0.03～0.05 part.
Preferably, in described biological organization sample preparation set liquid, also include being dehydrated liquid, in terms of volume parts, described de-
Water liquid include dehydrated alcohol 45～55 parts, the tert-butyl alcohol 15～25 parts, hexahydrotoluene 10～20 parts, acetone 6～9 parts and
Isopropanol 6～10 parts.
Preferably, in described biological organization sample preparation set liquid, described dehydration liquid includes dehydrated alcohol 50 parts, tertiary fourth
Alcohol 20 parts, hexahydrotoluene 15 parts, 7 parts of acetone and isopropanol 8 parts.
Preferably, in described biological organization sample preparation set liquid, also include waxdip liquid, in terms of parts by weight, described leaching
Wax liquid includes paraffin wax fully refined 70 parts, half paraffin wax fully refined 20 parts and 10 parts of refine honey lid Cera Flava and paraffin modification agent, institute
The ratio of the consumption and the gross weight of described paraffin wax fully refined, half paraffin wax fully refined and refine honey lid Cera Flava of stating paraffin modification agent is
3:1000。
Preferably, in described biological organization sample preparation set liquid, the preparation method of described waxdip liquid is particularly as follows: by waxdip
After the melting sources of liquid miscible uniformly, and at constant temperature 75 DEG C place 8～12h, room temperature cooling molding, to obtain described leaching
Wax liquid.
Preferably, in described biological organization sample preparation set liquid, described fixative also comprises natrium carbonicum calcinatum 3～5g/L
With trichloroacetic acid 6～8g/L.
Preferably, in described biological organization sample preparation set liquid, described fixative includes dehydrated alcohol 45 parts, nothing
Water methanol 25 parts, PEG400 15 parts and purified water 15 parts, the concentration of described natrium carbonicum calcinatum is 3g/L, described
The concentration of trichloroacetic acid is 7.5g/L.
Preferably, in described biological organization sample preparation set liquid, described fixative also includes 3 parts, 4 parts PBS of glacial acetic acid
Buffer and acetic acid 5 parts, the phosphate concentration in described PBS is 0.05～0.07mol/L.
Preferably, in described biological organization sample preparation set liquid, described transparent liquid also includes 5 parts of acetone and Oleum Terebinthinae 3
Part.
Preferably, in described biological organization sample preparation set liquid, described waxdip liquid also includes hexadecanoic acid 8 parts.
The present invention at least includes following beneficial effect:
The biological organization sample preparation set liquid of the present invention is nonhazardous environment-friendly type reagent.Meeting pathology routine diagnosis simultaneously,
Immunohistochemistry, the requirement of molecular biology experiment research can be met again.Conventional organization sample process has stopped tradition
Technology tissue samples is processed in often appearances make tissue samples contracting, firmly, split, crisp phenomenon, make cell and organizational structure more
Close to primary form, organizational structure is fixed intact, dehydration thoroughly；Due to this technology in immunohistochemical assay technology
Do not use formaldehyde, the most both avoided the toxic action of formaldehyde, eliminate again the formaldehyde covering to antigenic substance, it is possible to more preferably
Preservation antigen；In terms of molecular biology experiment and research, experiment proves that the tissue that this reagents series processes can well
Preserve the nucleic acid structure of cell, the beneficially separation of nucleic acid, extraction and correlation molecule pathological experiment.At this reagent
After reason tissue soft or hard appropriateness, both improve chipping qualities, the most significantly save microtome blade (refer mainly to hysteromyoma,
The sclerous tissues such as fibromyoma, skin), solve the problem that puzzlement pathology technicians's fatty tissue for many years is difficult to colouring.
Whole biological tissue preparation process, from the 13 of conventional art steps, tapers to 4 steps.This reagents series is to use
Difference in functionality the miscible compound product of the organic chemical reagent dissolved each other.Its raw material more than 97% is a class chemical reagent,
Remaining is two class chemical reagent, abandons three class chemical reagent.More single use formaldehyde is fixed, ethanol dehydration, and dimethylbenzene is transparent
Effect significantly improve.This reagents series feature is: low toxicity, degradable, pollution-free, minimizing discharge.By in conventional art
The poisonous and harmful of use, the formaldehyde of serious environment pollution, dimethylbenzene reagent are all abandoned, and eliminate consequent occupational environment
Pollute, from source, stop poisonous and harmful substance discharge, a large amount of minimizing reagent consumption.
Biological organization sample of the present invention preparation set pendular ring protects, formaldehydeless, without benzene, traditional preparation can be substituted completely and overlap liquid.With
Time, the preparation set safe and stable property of liquid of the present invention is good, and after configuration, Long time propertity keeps constant, and performance remains good, tool
There is good market value..
Part is embodied by the further advantage of the present invention, target and feature by description below, and part also will be by the present invention
Research and practice and be understood by the person skilled in the art.
Accompanying drawing explanation
Fig. 1 is the preparation set Skin slice prepared of liquid micro-utilizing the present invention in one of them embodiment of the present invention
Photo, amplification is 200 times.
Fig. 2 is the preparation set Skin slice prepared of liquid micro-utilizing the present invention in one of them embodiment of the present invention
Photo, amplification is 200 times.
Detailed description of the invention
The present invention is described in further detail below in conjunction with the accompanying drawings, to make those skilled in the art with reference to description word energy
Enough implement according to this.
Should be appreciated that used herein such as " have ", " comprising " and " including " term do not allot one or many
Other element individual or the existence of a combination thereof or interpolation.
The present invention provides a kind of biological organization sample preparation set liquid, including fixative and transparent liquid, in terms of volume parts,
Described fixative includes dehydrated alcohol 45～50 parts, absolute methanol 20～25 parts, PEG400 15 parts, pure
Change water 10～20 parts and surfactant 0.03～0.05 part；Described transparent liquid includes D-hesperidene 90 parts, isopropanol 10 parts
With surfactant 0.03～0.05 part.
In one of them embodiment of the present invention, as preferably, also include being dehydrated liquid, in terms of volume parts, described dehydration
Liquid includes dehydrated alcohol 45～55 parts, the tert-butyl alcohol 15～25 parts, hexahydrotoluene 10～20 parts, acetone 6～9 parts and different
Propanol 6～10 parts.
In one of them embodiment of the present invention, as preferably, described dehydration liquid includes dehydrated alcohol 50 parts, the tert-butyl alcohol
20 parts, hexahydrotoluene 15 parts, 7 parts of acetone and isopropanol 8 parts.
In one of them embodiment of the present invention, as preferably, also include waxdip liquid, in terms of parts by weight, described waxdip
Liquid includes paraffin wax fully refined 70 parts, half paraffin wax fully refined 20 parts and 10 parts of refine honey lid Cera Flava and paraffin modification agent, described
The consumption of paraffin modification agent with the ratio of the gross weight of described paraffin wax fully refined, half paraffin wax fully refined and refine honey lid Cera Flava is
3:1000。
In one of them embodiment of the present invention, as preferably, the preparation method of described waxdip liquid is particularly as follows: by waxdip liquid
Melting sources after miscible uniformly, and at constant temperature 75 DEG C place 8～12h, room temperature cooling molding, to obtain described waxdip
Liquid.
In one of them embodiment of the present invention, as preferably, described fixative also comprises natrium carbonicum calcinatum 3～5g/L
With trichloroacetic acid 6～8g/L.
In one of them embodiment of the present invention, as preferably, described fixative includes dehydrated alcohol 45 parts, anhydrous
Methanol 25 parts, PEG400 15 parts and purified water 15 parts, the concentration of described natrium carbonicum calcinatum is 3g/L, described three
Chloroacetic concentration is 7.5g/L.
In one of them embodiment of the present invention, as preferably, described fixative also includes 3 parts, 4 parts PBS of glacial acetic acid
Buffer and acetic acid 5 parts, the phosphate concentration in described PBS is 0.05～0.07mol/L.
In one of them embodiment of the present invention, as preferably, described transparent liquid also includes 5 parts of acetone and Oleum Terebinthinae 3 parts.
In one of them embodiment of the present invention, as preferably, described waxdip liquid also includes hexadecanoic acid 8 parts.
Embodiment 1
The biological organization sample preparation set liquid of the present invention, including:
Fixative: by volume, including dehydrated alcohol 45 parts, absolute methanol 25 parts, PEG400 15 parts, pure
Change 15 parts of water and 0.03 part of surfactant；And natrium carbonicum calcinatum 3g/L, trichloroacetic acid 7.5g/L, under room temperature by volume
Than miscible uniformly and get final product, finished product pH value 7～7.5.
Dehydration liquid: in terms of volume parts, dehydrated alcohol 50 parts, the tert-butyl alcohol 20 parts, hexahydrotoluene 15 parts, acetone 7
Part, isopropanol 8 parts, under room temperature the most miscible uniformly and get final product.
Transparent liquid: in terms of volume parts, D-hesperidene 90 parts, isopropanol 10 parts, surfactant 0.03～0.05 part, often
Under temperature the most miscible uniformly and get final product.
Waxdip: in terms of parts by weight, 56# paraffin wax fully refined 70 parts, 56# half paraffin wax fully refined 20 parts, refine honey lid honeybee
10 parts of wax；Paraffin modification agent 0.3g/kg, above-mentioned raw materials be placed in standing bucket in, after fusing miscible uniformly, at constant temperature 75 DEG C 8～
12h, room temperature cooling molding, fusing point: 56～58 DEG C.
Concrete operation method
For sake of clarity, tissue samples is processed the various functional reagent of part and is numbered.Particularly as follows: tissue fixative solution sets
Numbered first cylinder, tissue dewatering liquid is the second cylinder, and transparency of organization liquid is triplex, and tissue waxdip liquid is the 4th cylinder.Giving birth to
When fabric texture sample processes, biological organization sample is respectively put into the first cylinder successively to the 4th cylinder, and its operation requires as follows:
First cylinder soak time: 3～4h, the second cylinder soak time: 2.5～3h, triplex soak time: 2～3h；The
Four cylinder soak times: 3～4h.More than for biological tissue's sample tissue processing procedure, (above-mentioned soak time refers to normal structure
Block amasss 1.5 × 1.0 × 0.3；Soak time can vary in size according to different tissues, piece of tissue, makes the appropriate adjustments).
Embodiment 2
A kind of biological organization sample preparation set liquid, including fixative and transparent liquid, in terms of volume parts,
Described fixative includes dehydrated alcohol 50 parts, absolute methanol 20 parts, PEG400 15 parts, purified water 10 parts
With surfactant .05 part；With natrium carbonicum calcinatum 3g/L and trichloroacetic acid 6g/L
Dehydration liquid, in terms of volume parts, described dehydration liquid includes dehydrated alcohol 45 parts, the tert-butyl alcohol 15 parts, hexahydrotoluene
10 parts, 6 parts of acetone and isopropanol 6 parts.
Transparent liquid includes D-hesperidene 90 parts, isopropanol 10 parts and surfactant 0.03～0.05 part.
Waxdip liquid, in terms of parts by weight, described waxdip liquid includes paraffin wax fully refined 70 parts, half paraffin wax fully refined 20 parts and essence
10 parts of refined honey lid Cera Flava and paraffin modification agent, the consumption of described paraffin modification agent and described paraffin wax fully refined, half full refining stone
The ratio of the gross weight of wax and refine honey lid Cera Flava is 3:1000.The preparation method of described waxdip liquid is particularly as follows: former by waxdip liquid
Material fusing after miscible uniformly, and at constant temperature 75 DEG C place 8～12h, room temperature cooling molding, to obtain described waxdip liquid.
Embodiment 3
A kind of biological organization sample preparation set liquid, comprises:
Fixative, in terms of volume parts, including dehydrated alcohol 47 parts, absolute methanol 22 parts, PEG400 15 parts,
Purified water 20 parts and 0.04 part of surfactant；And natrium carbonicum calcinatum 5g/L and trichloroacetic acid 8g/L.
Dehydration liquid, in terms of volume parts, including dehydrated alcohol 55 parts, the tert-butyl alcohol 25 parts, hexahydrotoluene 20 parts, third
Ketone 9 parts and isopropanol 10 parts.
Transparent liquid, in terms of volume parts, including D-hesperidene 90 parts, isopropanol 10 parts and 0.04 part of surfactant.
Waxdip liquid, in terms of parts by weight, described waxdip liquid includes paraffin wax fully refined 70 parts, half paraffin wax fully refined 20 parts and essence
10 parts of refined honey lid Cera Flava and paraffin modification agent, the consumption of described paraffin modification agent and described paraffin wax fully refined, half full refining stone
The ratio of the gross weight of wax and refine honey lid Cera Flava is 3:1000.The preparation method of waxdip liquid is particularly as follows: melt the raw material of waxdip liquid
After change miscible uniformly, and at constant temperature 75 DEG C place 8～12h, room temperature cooling molding, to obtain described waxdip liquid.
Embodiment 4
A kind of biological organization sample preparation set liquid, comprises:
Fixative, in terms of volume parts, including dehydrated alcohol 47 parts, absolute methanol 22 parts, PEG400 15 parts,
3 parts, 4 parts PBS of glacial acetic acid, acetic acid 5 parts, purified water 20 parts and 0.04 part of surfactant；And anhydrous carbon
Acid sodium 5g/L and trichloroacetic acid 8g/L.Phosphate concentration in described PBS is 0.05～0.07mol/L.The present invention
Fixative can quickly biological organization sample be realized fixing, the active substance of biological organization sample is affected the least.
Dehydration liquid, in terms of volume parts, including dehydrated alcohol 55 parts, the tert-butyl alcohol 25 parts, hexahydrotoluene 20 parts, third
Ketone 9 parts and isopropanol 10 parts.
Transparent liquid, in terms of volume parts, including D-hesperidene 90 parts, isopropanol 10 parts, 5 parts of acetone, Oleum Terebinthinae 3 parts
With 0.04 part of surfactant.In transparent liquid, whole components are nontoxic chemical reagent, jointly to biological tissue's sample
Originally transparent effect is played.
Waxdip liquid, in terms of parts by weight, described waxdip liquid includes paraffin wax fully refined 70 parts, half paraffin wax fully refined 20 parts and essence
10 parts of refined honey lid Cera Flava, hexadecanoic acid 8 parts and paraffin modification agent, the consumption of described paraffin modification agent and described full refining stone
The ratio of the gross weight of wax, half paraffin wax fully refined and refine honey lid Cera Flava is 3:1000.The preparation method of waxdip liquid is particularly as follows: incite somebody to action
After the melting sources of waxdip liquid miscible uniformly, and at constant temperature 75 DEG C place 8～12h, room temperature cooling molding, to obtain
State waxdip liquid.
Module number described herein and treatment scale are used to the explanation of the simplification present invention.To the fixative of the present invention, de-
Water liquid, transparent liquid and the application of waxdip liquid, modifications and variations will be readily apparent to persons skilled in the art.
Although embodiment of the present invention are disclosed as above, but it is not restricted in description and embodiment listed fortune
With, it can be applied to various applicable the field of the invention completely, for those skilled in the art, and can be easily
Realizing other amendment, therefore under the general concept limited without departing substantially from claim and equivalency range, the present invention does not limit
In specific details with shown here as the legend with description.
Claims (10)
1. a biological organization sample preparation set liquid, including fixative and transparent liquid, it is characterised in that in terms of volume parts,
Described fixative includes dehydrated alcohol 45～50 parts, absolute methanol 20～25 parts, PEG400 15 parts, pure
Change water 10～20 parts and surfactant 0.03～0.05 part；
Described transparent liquid includes D-hesperidene 90 parts, isopropanol 10 parts and surfactant 0.03～0.05 part.
2. biological organization sample preparation set liquid as claimed in claim 1, it is characterised in that also include being dehydrated liquid, with body
Long-pending number meter, described dehydration liquid includes dehydrated alcohol 45～55 parts, the tert-butyl alcohol 15～25 parts, hexahydrotoluene 10～20
Part, acetone 6～9 parts and isopropanol 6～10 parts.
3. biological organization sample preparation set liquid as claimed in claim 2, it is characterised in that described dehydration liquid includes anhydrous
Ethanol 50 parts, the tert-butyl alcohol 20 parts, hexahydrotoluene 15 parts, 7 parts of acetone and isopropanol 8 parts.
4. biological organization sample preparation set liquid as claimed in claim 1, it is characterised in that also include waxdip liquid, with weight
Amount number meter, described waxdip liquid includes paraffin wax fully refined 70 parts, half paraffin wax fully refined 20 parts and 10 parts of refine honey lid Cera Flava
With paraffin modification agent, the consumption of described paraffin modification agent and described paraffin wax fully refined, half paraffin wax fully refined and refine honey lid Cera Flava
The ratio of gross weight be 3:1000.
5. biological organization sample preparation set liquid as claimed in claim 1, it is characterised in that the preparation side of described waxdip liquid
Method particularly as follows: by after the melting sources of waxdip liquid miscible uniformly, and at constant temperature 75 DEG C place 8～12h, room temperature is cooled to
Type, to obtain described waxdip liquid.
6. biological organization sample preparation set liquid as claimed in claim 1, it is characterised in that also comprise in described fixative
Natrium carbonicum calcinatum 3～5g/L and trichloroacetic acid 6～8g/L.
7. biological organization sample preparation set liquid as claimed in claim 6, it is characterised in that described fixative includes nothing
Water-ethanol 45 parts, absolute methanol 25 parts, PEG400 15 parts and purified water 15 parts, described natrium carbonicum calcinatum dense
Degree is 3g/L, and the concentration of described trichloroacetic acid is 7.5g/L.
8. biological organization sample preparation set liquid as claimed in claim 1, it is characterised in that described fixative also includes ice
3 parts, 4 parts PBS of acetic acid and acetic acid 5 parts, the phosphate concentration in described PBS is 0.05～0.07mol/L.
9. biological organization sample preparation set liquid as claimed in claim 1, it is characterised in that described transparent liquid also includes third
Ketone 5 parts and Oleum Terebinthinae 3 parts.
10. biological organization sample preparation set liquid as claimed in claim 4, it is characterised in that described waxdip liquid also includes ten
Six alkanoic acids 8 parts.
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| CN201610319967.5A CN105823665A (en) | 2016-05-13 | 2016-05-13 | Biological tissue sample preparation sleeve solution |
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| CN201610319967.5A CN105823665A (en) | 2016-05-13 | 2016-05-13 | Biological tissue sample preparation sleeve solution |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108918213A (en) * | 2018-05-16 | 2018-11-30 | 浙江思格医疗科技有限公司 | A kind of palmitin fixer |
| CN110426259A (en) * | 2019-08-14 | 2019-11-08 | 武汉赛维尔生物科技有限公司 | Polyethylene glycol is used for the application of animal tissue sections grease dyeing |
| CN111795879A (en) * | 2020-07-14 | 2020-10-20 | 青岛大学附属医院 | Application of pathological specimen manufactured by dehydration set in digital evaluation system |
| CN112067382A (en) * | 2020-08-07 | 2020-12-11 | 佛山科学技术学院 | Preparation method of small intestine slices |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108918213A (en) * | 2018-05-16 | 2018-11-30 | 浙江思格医疗科技有限公司 | A kind of palmitin fixer |
| CN110426259A (en) * | 2019-08-14 | 2019-11-08 | 武汉赛维尔生物科技有限公司 | Polyethylene glycol is used for the application of animal tissue sections grease dyeing |
| CN111795879A (en) * | 2020-07-14 | 2020-10-20 | 青岛大学附属医院 | Application of pathological specimen manufactured by dehydration set in digital evaluation system |
| CN112067382A (en) * | 2020-08-07 | 2020-12-11 | 佛山科学技术学院 | Preparation method of small intestine slices |
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