NO742274L - - Google Patents
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- Publication number
- NO742274L NO742274L NO742274A NO742274A NO742274L NO 742274 L NO742274 L NO 742274L NO 742274 A NO742274 A NO 742274A NO 742274 A NO742274 A NO 742274A NO 742274 L NO742274 L NO 742274L
- Authority
- NO
- Norway
- Prior art keywords
- approx
- tissue
- solution
- trichloroacetic acid
- hematoxylin
- Prior art date
Links
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims description 42
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 claims description 40
- 239000000243 solution Substances 0.000 claims description 27
- JIAARYAFYJHUJI-UHFFFAOYSA-L zinc dichloride Chemical compound [Cl-].[Cl-].[Zn+2] JIAARYAFYJHUJI-UHFFFAOYSA-L 0.000 claims description 26
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 24
- 238000000034 method Methods 0.000 claims description 20
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 claims description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- 239000011592 zinc chloride Substances 0.000 claims description 13
- 235000005074 zinc chloride Nutrition 0.000 claims description 13
- 238000007654 immersion Methods 0.000 claims description 8
- 239000000203 mixture Substances 0.000 claims description 6
- 238000010186 staining Methods 0.000 claims description 5
- AXDJCCTWPBKUKL-UHFFFAOYSA-N 4-[(4-aminophenyl)-(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methyl]aniline;hydron;chloride Chemical group Cl.C1=CC(=N)C(C)=CC1=C(C=1C=CC(N)=CC=1)C1=CC=C(N)C=C1 AXDJCCTWPBKUKL-UHFFFAOYSA-N 0.000 claims description 4
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical group [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 claims description 4
- XGZVUEUWXADBQD-UHFFFAOYSA-L lithium carbonate Chemical compound [Li+].[Li+].[O-]C([O-])=O XGZVUEUWXADBQD-UHFFFAOYSA-L 0.000 claims description 4
- 229910052808 lithium carbonate Inorganic materials 0.000 claims description 4
- 229940037003 alum Drugs 0.000 claims description 3
- 239000007864 aqueous solution Substances 0.000 claims description 3
- 239000000994 contrast dye Substances 0.000 claims description 2
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 claims description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 19
- 210000001519 tissue Anatomy 0.000 description 18
- 238000002360 preparation method Methods 0.000 description 8
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 7
- 239000012153 distilled water Substances 0.000 description 6
- 239000000834 fixative Substances 0.000 description 5
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 4
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 239000008096 xylene Substances 0.000 description 4
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 3
- 239000000975 dye Substances 0.000 description 3
- 235000011187 glycerol Nutrition 0.000 description 3
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- 229960000583 acetic acid Drugs 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 239000012362 glacial acetic acid Substances 0.000 description 2
- 229910000474 mercury oxide Inorganic materials 0.000 description 2
- UKWHYYKOEPRTIC-UHFFFAOYSA-N mercury(ii) oxide Chemical compound [Hg]=O UKWHYYKOEPRTIC-UHFFFAOYSA-N 0.000 description 2
- OXNIZHLAWKMVMX-UHFFFAOYSA-N picric acid Chemical compound OC1=C([N+]([O-])=O)C=C([N+]([O-])=O)C=C1[N+]([O-])=O OXNIZHLAWKMVMX-UHFFFAOYSA-N 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 2
- 239000012192 staining solution Substances 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- WZUVPPKBWHMQCE-XJKSGUPXSA-N (+)-haematoxylin Chemical compound C12=CC(O)=C(O)C=C2C[C@]2(O)[C@H]1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-XJKSGUPXSA-N 0.000 description 1
- HZLHRDBTVSZCBS-GHTYLULLSA-N 4-[(z)-(4-aminophenyl)-(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methyl]-2-methylaniline;hydrochloride Chemical compound Cl.C1=CC(=N)C(C)=C\C1=C(C=1C=C(C)C(N)=CC=1)\C1=CC=C(N)C=C1 HZLHRDBTVSZCBS-GHTYLULLSA-N 0.000 description 1
- WZUKKIPWIPZMAS-UHFFFAOYSA-K Ammonium alum Chemical compound [NH4+].O.O.O.O.O.O.O.O.O.O.O.O.[Al+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O WZUKKIPWIPZMAS-UHFFFAOYSA-K 0.000 description 1
- 208000019901 Anxiety disease Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- RZUBARUFLYGOGC-MTHOTQAESA-L acid fuchsin Chemical compound [Na+].[Na+].[O-]S(=O)(=O)C1=C(N)C(C)=CC(C(=C\2C=C(C(=[NH2+])C=C/2)S([O-])(=O)=O)\C=2C=C(C(N)=CC=2)S([O-])(=O)=O)=C1 RZUBARUFLYGOGC-MTHOTQAESA-L 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 239000001063 aluminium ammonium sulphate Substances 0.000 description 1
- 235000011124 aluminium ammonium sulphate Nutrition 0.000 description 1
- LCQXXBOSCBRNNT-UHFFFAOYSA-K ammonium aluminium sulfate Chemical compound [NH4+].[Al+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O LCQXXBOSCBRNNT-UHFFFAOYSA-K 0.000 description 1
- 230000036506 anxiety Effects 0.000 description 1
- 229940052223 basic fuchsin Drugs 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000012631 diagnostic technique Methods 0.000 description 1
- 238000007598 dipping method Methods 0.000 description 1
- 230000009189 diving Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- -1 for 4 seconds Chemical compound 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000007431 microscopic evaluation Methods 0.000 description 1
- IPSIPYMEZZPCPY-UHFFFAOYSA-N new fuchsin Chemical compound [Cl-].C1=CC(=[NH2+])C(C)=CC1=C(C=1C=C(C)C(N)=CC=1)C1=CC=C(N)C(C)=C1 IPSIPYMEZZPCPY-UHFFFAOYSA-N 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- AFAIELJLZYUNPW-UHFFFAOYSA-N pararosaniline free base Chemical compound C1=CC(N)=CC=C1C(C=1C=CC(N)=CC=1)=C1C=CC(=N)C=C1 AFAIELJLZYUNPW-UHFFFAOYSA-N 0.000 description 1
- GRLPQNLYRHEGIJ-UHFFFAOYSA-J potassium aluminium sulfate Chemical compound [Al+3].[K+].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O GRLPQNLYRHEGIJ-UHFFFAOYSA-J 0.000 description 1
- 238000011470 radical surgery Methods 0.000 description 1
- 239000012047 saturated solution Substances 0.000 description 1
- 238000007447 staining method Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- 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
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/0231—Chemically defined matrices, e.g. alginate gels, for immobilising, holding or storing cells, tissue or organs for preservation purposes; Chemically altering or fixing cells, tissue or organs, e.g. by cross-linking, for preservation purposes
-
- 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
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
- G01N2001/305—Fixative compositions
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Dispersion Chemistry (AREA)
- Dentistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Sampling And Sample Adjustment (AREA)
Description
Fremgangsmåte for farvning av vev. Procedure for staining tissue.
Nærværende oppfinnelse vedrorer en histologisk farvnings-metode for vevprover for mikroskopisk undersokelse. Den selektive farvning av vevceller både for å identifisere forskjellige deler av cellen og å skjelne forskjellige typer celler er en velkjent diagnostisk teknikk. Egnede fremgangsmåter for å gi reproduserbare resultater er ofte tidskrevende og besværlige. Ved en typisk ryogen anvendelse folges to hoved-arbeidsmåter. Vevet fryses hurtig og deles i snitt. Disse snitt fikseres f.eks. i formaldehyd eller alkohol, farves med en enkel farve, dekkes med dekkglass og undersokes. Dette The present invention relates to a histological staining method for tissue samples for microscopic examination. The selective staining of tissue cells both to identify different parts of the cell and to distinguish different types of cells is a well-known diagnostic technique. Suitable methods for producing reproducible results are often time-consuming and cumbersome. In a typical ryogenic application, two main working methods are followed. The tissue is quickly frozen and cut into sections. These sections are fixed e.g. in formaldehyde or alcohol, stained with a simple dye, covered with a glass coverslip and examined. This
preparat er ikke permanent og har en begrenset levetid påpreparation is not permanent and has a limited lifespan
bare noen få timer. Skjont den tillater en mer eller mindre hurtig undersokelse av vevet, har arbeidsmetoden flere ulemper, av hvilke det forskjellige utseendet for prover fra vanlig mikroskopiske preparater er den storste og hvilket oker vanskeligheten med å oppnå en riktig fortolkning. just a few hours. Although it allows a more or less rapid examination of the tissue, the working method has several disadvantages, of which the different appearance of samples from ordinary microscopic preparations is the biggest and which increases the difficulty in achieving a correct interpretation.
For å kunne utfore en mer definitiv mikroskopisk analyse ogIn order to perform a more definitive microscopic analysis and
et permanent preparat av det originale vev folges en annen arbeidsmåte. Her tines det frosne vev, fikseres, dehydratiseres, behandles med organiske opplosningsmidler, legges ned i parafin og snitt fremstilles. Disse snitt avparafineres i tur og orden, hydratiseres, farves, dehydratiseres og dekkes med dekkglass. Dette gir et permanent mikroskopisk preparat av materialet opprinnelig undersokt ved den temporære kryogene metode. a permanent preparation of the original tissue follows a different method of working. Here the frozen tissue is thawed, fixed, dehydrated, treated with organic solvents, embedded in paraffin and sections are prepared. These sections are deparaffinized in turn, hydrated, stained, dehydrated and covered with coverslips. This provides a permanent microscopic preparation of the material originally examined by the temporary cryogenic method.
Som det vil bemerkes er denne arbeidsmåte meget lang og besværlig og dessuten er provene ikke en noyaktig gjengivelse av det originale vev. I tillegg til den mulige skade som er forårsaket av iskrystalldannelse, tapes noe vev i lopet av den kjemiske og fysiske behandling. As will be noted, this method of working is very long and cumbersome and, moreover, the sample is not an accurate reproduction of the original tissue. In addition to the possible damage caused by ice crystal formation, some tissue is lost in the course of the chemical and physical treatment.
Samspillet mellom disse to arbeidsmåter, og deres ledsagende ulemper, kan lettest f .eks. sees i tilfellet av antatt ondartet-het. Typisk vil en vevprove f j ernet fra en pasient i lopet av en under-sokende operasjon gjores til gjenstand for den forste arbeidsmåte for å tillate kirurgen å ta en umiddelbar avgjorelse vedrorende naturen og graden av radikal operasjon. Fremstilling av en permanent mikroskopisk prove for å få bekreftelse på diagnosen kan kreve så meget som 24 timer eller mer, hvilket klart begrenser den mikroskopiske undersokelse av vev som fjernes under operasjon til de mindre definitive temporære preparater da pasienten ikke kan holdes i operasjonssalen i den lange periode som kreves for en permanent fremstilling. Dessuten kjennes ikke endelige resultater fra vevstudier på flere dager og derved okeseller forlenges den naturlige uro hos pasient og familie. Da de prover ved hvilke diagnosen var basert ofte ikke er egnet for oppbevaring, må besværlige preserverings-trinn tas hvis den medisinske tilstand eller hospitale politikk The interaction between these two ways of working, and their accompanying disadvantages, can most easily e.g. seen in the case of presumed malignancy. Typically, a tissue sample removed from a patient in the course of an exploratory operation will be the subject of the first procedure to allow the surgeon to make an immediate decision regarding the nature and degree of radical surgery. Preparation of a permanent microscopic specimen to confirm the diagnosis may require as much as 24 hours or more, which clearly limits the microscopic examination of tissue removed during surgery to the less definitive temporary preparations as the patient cannot be kept in the operating theater for the long period required for a permanent manufacture. In addition, final results from tissue studies are not known for several days, thereby prolonging the natural anxiety of the patient and family. As the samples on which the diagnosis was based are often not suitable for preservation, difficult preservation steps must be taken if the medical condition or hospital policy
nodvendiggjor oppbevaring av proven i mer enn noen få dager.require storage of the sample for more than a few days.
I U.S. patentsoknad Ser. No. 338 732 beskrives et nyttIn the U.S. patent application Ser. No. 338 732 a new one is described
histologisk fiksativ som består av en opplosning av trikloreddiksyre, sinkklorid og formaldehyd i en vandig lavere alkanol. Dette fiksativ forårsaker minimal skade på .vevcellen, histological fixative consisting of a solution of trichloroacetic acid, zinc chloride and formaldehyde in an aqueous lower alkanol. This fixative causes minimal damage to the tissue cell,
er lettvint i bruk og kan brukes i forbindelse med en rekke varianter av vevtyper. Det er nå blitt funnet at dette fiksativ kan brukes i en meget forbedret metode ved å farve vevtyper. Den totale farvningsprosess kan utfores i lopet av minutter og de resulterende farvede prover oppviser tilstrek-kelig permanens for å tillate oppbevaring av disse i måneder uten spesielle foranstaltninger. is easy to use and can be used in connection with a number of variants of tissue types. It has now been found that this fixative can be used in a much improved method by staining tissue types. The total dyeing process can be carried out in a matter of minutes and the resulting dyed samples exhibit sufficient permanence to allow their storage for months without special measures.
Prover som skal farves fryses hurtig ved en av de velkjente metoder. Fra det frosne vev fremstilles derpå snitt med en vanlig mikrotom og snittene senkes ned i en fikseringsopplosning som består av en vandig lavere alkanolisk opplosning av trikloreddiksyre, sinkklorid og formaldehyd. Snittet er allerede selvfolgelig tint opp for denne neddykking men fikseres på sekunder. En typisk periode for neddykking er 5 sekunder og skjont denne periode kan forlenges, så blir resul-tatet ikke mer fordelaktig ved en lenger neddykking. Samples to be stained are frozen quickly by one of the well-known methods. Sections are then made from the frozen tissue with an ordinary microtome and the sections are immersed in a fixing solution consisting of an aqueous lower alkanol solution of trichloroacetic acid, zinc chloride and formaldehyde. The cut is obviously already thawed for this dive but is fixed in seconds. A typical period for submersion is 5 seconds and although this period can be extended, the result is not more beneficial with a longer submersion.
Snittet dykkes deretter ned i en vandig opplosning avThe section is then immersed in an aqueous solution of
hematoxylin. Hematoxylin er en velkjent farve som kjemisk er 7,llb-dihydrobenz[b]indeno[l,2-d]pyran-3,4,6a,9,10(6H)-pentol. Dette trinn av fremgangsmåten kan utfores i ett trinn eller hematoxylin. Hematoxylin is a well-known dye which is chemically 7,11b-dihydrobenz[b]indeno[1,2-d]pyran-3,4,6a,9,10(6H)-pentol. This step of the procedure can be performed in one step or
to trinn. Ved en utforelsesform senkes snittet ned i opplosningen av hematoxylin inntil farvningen er fullstendig, hvilket vanligvis er fra 10 til 30 sekunder. Ved en annen utforelsesform senkes proven ned for et tidsrom i en forste opplosning av hematoxylin, fjernes og renses med vann og senkes ned i en annen opplosning av hematoxylin. Ved et foretrukket trekk ved denne annen utforelsesform forutgås nedsenkningen av proven i den annen opplosning av hematoxylin ved en kort nedsenkning i en opplosning av litiumkarbonat som virker som en aksellerator. Således dykkes f.eks. snittet fra two steps. In one embodiment, the section is immersed in the solution of hematoxylin until the staining is complete, which is usually from 10 to 30 seconds. In another embodiment, the sample is immersed for a period of time in a first solution of hematoxylin, removed and cleaned with water and immersed in another solution of hematoxylin. In a preferred feature of this second embodiment, the immersion of the sample in the second solution of hematoxylin is preceded by a short immersion in a solution of lithium carbonate which acts as an accelerator. Thus, e.g. diving cut from
fikseringsopplosningen ned i en forste hematoxylinopplosning i ca. 15 sekunder, skylles ved å dyppe den ned i vann, senkes kort ned i en opplosning av litiumkarbonat, f.eks. i 4 sekunder, the fixing solution into a first hematoxylin solution for approx. 15 seconds, rinsed by dipping it in water, briefly immersed in a solution of lithium carbonate, e.g. for 4 seconds,
og senkes igjen ned i en opplosning av hematoxylin i ca. 5 sekunder. Etter dette er proven fiksert og cellekjernene farvet med en blålig purpurfarve. Proven kan derpå prepareres for montering ved vasking og torking, det siste utfores på and immersed again in a solution of hematoxylin for approx. 5 seconds. After this, the sample is fixed and the cell nuclei stained with a bluish purple colour. The sample can then be prepared for assembly by washing and drying, the latter being carried out on
vanlig måte ved etterfølgende neddykking i progressivt mer hydrofile organiske media, f.eks. 95% etanol, absolutt.etanol og xylen. usual way by subsequent immersion in progressively more hydrophilic organic media, e.g. 95% ethanol, absolute ethanol and xylene.
Hvis onsket kan proven også kontrafarves ved bruk av deIf desired, the sample can also be counterstained using de
velkjente differensierende farver som f.eks. eosin, en fuchsin slik som rosanilin, pararosanilin, magenta II, magenta III well-known differentiating colors such as eosin, a fuchsin such as rosaniline, pararosaniline, magenta II, magenta III
eller sur fuchsin; pikrinsyre eller lignende.or acid fuchsin; picric acid or the like.
Som angitt foran består fiksativet av en opplosning av trikloreddiksyre, sinkklorid og formaldehyd i en vandig lavere alkanol. Den lavere alkanol kan være enhver at de velkjente alkoholer As indicated above, the fixative consists of a solution of trichloroacetic acid, zinc chloride and formaldehyde in an aqueous lower alkanol. The lower alkanol can be any of the well-known alcohols
som har fra 1 til 4 karbonatomer som for eksempel metanol, etanol, n-propanol, isopropanol, n-butanol og de forskjellige forgrenede butanoler. Metanol er meget tilfredsstillende og foretrukket ut fra okonomiske betraktninger og opplosningsevne.. Den vandige lavere alkanol vil ha et volumforhold mellom alkohol og vann på ca. 1:1 til 1:3.- Et foretrukket forhold er ca. which have from 1 to 4 carbon atoms such as methanol, ethanol, n-propanol, isopropanol, n-butanol and the various branched butanols. Methanol is very satisfactory and preferred based on economic considerations and solubility. The aqueous lower alkanol will have a volume ratio between alcohol and water of approx. 1:1 to 1:3.- A preferred ratio is approx.
1:1.1 til 1:1.5. Opplost i den vandige alkohol er en blanding av trikloreddiksyre, sinkklorid og formaldehyd. Et forhold mellom sinkklorid og trikloreddiksyre, på vektsbasis, er ca. 1:1.1 to 1:1.5. Dissolved in the aqueous alcohol is a mixture of trichloroacetic acid, zinc chloride and formaldehyde. A ratio between zinc chloride and trichloroacetic acid, on a weight basis, is approx.
fra 2:1 til 4:1. Et forhold på 3:1 er meget tilfredsstillende Forholdet mellom formaldehyd og trikloreddiksyre er fra ca. from 2:1 to 4:1. A ratio of 3:1 is very satisfactory. The ratio between formaldehyde and trichloroacetic acid is from approx.
8:1 til 10:1, et typisk forhold er 9:1.8:1 to 10:1, a typical ratio is 9:1.
En foretrukket endelig konsentrasjon er ca. 10% vekt/volum av trikloreddiksyre, sinkklorid og formaldehyd i den vandig lavere alkanol. A preferred final concentration is approx. 10% w/v of trichloroacetic acid, zinc chloride and formaldehyde in the aqueous lower alkanol.
Hematoxylinet kan foreligge i enhver av de vanlige former.Hematoxylin can be present in any of the usual forms.
Disse omfatter Harris' Hematoxylin, Mayer's Haemalum,These include Harris' Hematoxylin, Mayer's Haemalum,
Erlich's Hematoxylin, alunhematoxylinet beskrevet av Lie et al., Mayo Clinic Proceedings, 46 319 (May 1971), og lignende. Erlich's Hematoxylin, the alum hematoxylin described by Lie et al., Mayo Clinic Proceedings, 46 319 (May 1971), and the like.
Et særlig anvendelig preparat er ett som inneholder hematoxylin, gult mercuryoksyd og aluminium-ammoniumsulfat i vandig glycerin , og denne reagens er generelt kjent som alun-hematoxylin. A particularly useful preparation is one containing hematoxylin, yellow mercury oxide and aluminum ammonium sulphate in aqueous glycerin, and this reagent is generally known as alum hematoxylin.
PreparaterPreparations
A. FikserinqsopplosninqA. Fixing solutions
400 g sinkklorid blandes med 120g trikloreddiksyre og 2.64 1400 g zinc chloride is mixed with 120 g trichloroacetic acid and 2.64 1
37% formaldehyd (tilsvarende 1055 g 100% formaldehyd) i 7.3 137% formaldehyde (equivalent to 1055 g 100% formaldehyde) in 7.3 1
95% metanol (tilsvarende 7 1 100% metanol) og 7 liter vann.95% methanol (equivalent to 7 1 100% methanol) and 7 liters of water.
En svak eksotermisk reaksjon inntreffer og etter at opplosningen har gjenvunnet romtemperatur, filtreres den og fylles på A slight exothermic reaction occurs and after the solution has regained room temperature it is filtered and replenished
beholder. Denne opplosning som er ferdig for bruk som et vevfiksativ, tilsvarer en 9,8 vekts%/volum opplosning av trikloreddiksyre, sinkklorid og formaldehyd, hvor vektsforhoIdet • av sinkklorid til trikloreddiksyre er ca. 3.33:1 og vektsforholdet mellom formaldehyd og trikloreddiksyre er ca. 9:1, container. This solution, which is ready for use as a tissue fixative, corresponds to a 9.8% by weight/volume solution of trichloroacetic acid, zinc chloride and formaldehyde, where the weight ratio • of zinc chloride to trichloroacetic acid is approx. 3.33:1 and the weight ratio between formaldehyde and trichloroacetic acid is approx. 9:1,
i vandig metanol hvor volumforholdet mellom alkanol og totalt vann (inklusive det som tilsettes separat, som inneholdes i den 95%'ige metanol og det som er inneholdt i den 37%'ige formaldehyd) er ca. 1:1.3. in aqueous methanol where the volume ratio between alkanol and total water (including what is added separately, which is contained in the 95% methanol and what is contained in the 37% formaldehyde) is approx. 1:1.3.
B. ( I) farqeopplosningB. ( I) farce solution
Til en opplosning av 0.67 g hematoxylin i 33 ml absoluttTo a solution of 0.67 g of hematoxylin in 33 ml of absolute
etanol tilsettes 33 ml destillert vann, 33 ml glycerol, 3.3 ml iseddik og et overskudd av kalium-aluminium-sulfat. Blandingen rystes godt om og får henstå i flere uker, hvoretter den er ferdig for bruk. ethanol, 33 ml of distilled water, 33 ml of glycerol, 3.3 ml of glacial acetic acid and an excess of potassium aluminum sulphate are added. The mixture is shaken well and allowed to stand for several weeks, after which it is ready for use.
B. ( 2) farqeopplosningB. ( 2) farce resolution
1 gram hematoxylin, 0.5 g gult mercuryoksyd og 12 g aluminium-ammonium-sulfat blandes med 140 ml destillert vann. Denne blanding kokes i 10 minutter, avkjoles og justeres til sitt opprinnelige volum med destillert vann. 60 milliliter glycerin og 8 ml iseddiksyre tilsettes og blandingen filtreres deretter. 1 gram of haematoxylin, 0.5 g of yellow mercury oxide and 12 g of aluminium-ammonium sulphate are mixed with 140 ml of distilled water. This mixture is boiled for 10 minutes, cooled and adjusted to its original volume with distilled water. 60 milliliters of glycerin and 8 ml of glacial acetic acid are added and the mixture is then filtered.
C. kontrastfarveopplosninqC. contrast dye solution
Denne kan kjopes som sådan og omfatter vandige opplosninger av eosin, basisk fuchsin, pikrinsyre og lignende. This can be bought as such and includes aqueous solutions of eosin, basic fuchsin, picric acid and the like.
EKSEMPEL 1EXAMPLE 1
Frosne vevprover deles opp i snitt med en mikrotom til enFrozen tissue samples are sectioned with a microtome to one
tykkelse på ca. 5ju. De individuelle snitt dyppes ned i fiks-seringsopplosningen i tilnærmelsesvis 5 sekunder og deretter i farveopplosningen B(2) i tilnærmet 15 sekunder. Etter ned-dypping i destillert vann i ca. 3 sekunder, dyppes snittene ned i en mettet opplosning av litiumkarbonat i omtrent 4 sekunder og dyppes imidlertid deretter igjen ned i farveopplosning B(2) thickness of approx. 5ju. The individual sections are immersed in the fixing solution for approximately 5 seconds and then in the staining solution B(2) for approximately 15 seconds. After immersion in distilled water for approx. 3 seconds, the sections are immersed in a saturated solution of lithium carbonate for approximately 4 seconds and then, however, immersed again in dye solution B(2)
i ca. 5 sekunder. Det fikserte og farvede snitt renses derpåfor about. 5 seconds. The fixed and stained section is then cleaned
i destillert vann i 3 sekunder og senkes derpå i rekkefolge ned i in distilled water for 3 seconds and then successively lowered into
.95% etanol, absolutt etanol, xylen nr. 1 og xylen nr. 2,.95% ethanol, absolute ethanol, xylene No. 1 and xylene No. 2,
hver neddykking varer i ca. 2 sekunder, hvoretter snittet dekkes med et dekkglass. each dive lasts approx. 2 seconds, after which the incision is covered with a coverslip.
EKSEMPEL 2EXAMPLE 2
Et kontra stfar vet snitt oppnås ifolge. fremgangsmåten i eksempel 1 ved å rense proven med destillert vann i 3 sekunder etter at den er fjernet fra den annen neddykking iB(2) farvnings-opplosningen, senke den ned i 95% etanol i 2 sekunder og derpå dyppe denne ned i en standard eosinopplosning i cai 15 sekunder. Det således differensierte snitt senkes derpå i rekkefolge ned i 95% etanol, absolutt etanol og xylen som beskrevet i eksempel I. A contrasting section is obtained as a result. the procedure in example 1 by cleaning the sample with distilled water for 3 seconds after removing it from the second immersion in the B(2) staining solution, immersing it in 95% ethanol for 2 seconds and then immersing it in a standard eosin solution for about 15 seconds. The thus differentiated section is then immersed in order in 95% ethanol, absolute ethanol and xylene as described in example I.
Claims (7)
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US37327673A | 1973-06-25 | 1973-06-25 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| NO742274L true NO742274L (en) | 1975-01-20 |
Family
ID=23471724
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| NO742274A NO742274L (en) | 1973-06-25 | 1974-06-21 |
Country Status (16)
| Country | Link |
|---|---|
| JP (1) | JPS5049026A (en) |
| AU (1) | AU7049174A (en) |
| BE (1) | BE816786R (en) |
| BR (1) | BR7405213D0 (en) |
| CA (1) | CA1037366A (en) |
| DD (1) | DD112519A5 (en) |
| DE (1) | DE2429647A1 (en) |
| DK (1) | DK338574A (en) |
| ES (1) | ES427568A1 (en) |
| FI (1) | FI193074A (en) |
| FR (1) | FR2234558B2 (en) |
| HU (1) | HU168582B (en) |
| NL (1) | NL7408556A (en) |
| NO (1) | NO742274L (en) |
| SE (1) | SE7407899L (en) |
| ZA (1) | ZA743995B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH068818B2 (en) * | 1987-11-27 | 1994-02-02 | サクラ精機株式会社 | Fixative for biopsy |
| EP1605244A1 (en) * | 2004-06-09 | 2005-12-14 | Boon, Mathilde Elisabeth | Fixative composition |
| CN110823666A (en) * | 2019-11-27 | 2020-02-21 | 李雄 | Application of hematoxylin in preparation of staining agent for reducing color fading of pathological conventional section after staining |
-
1974
- 1974-06-14 SE SE7407899A patent/SE7407899L/ not_active Application Discontinuation
- 1974-06-20 DE DE2429647A patent/DE2429647A1/en active Pending
- 1974-06-21 CA CA203,034A patent/CA1037366A/en not_active Expired
- 1974-06-21 ZA ZA00743995A patent/ZA743995B/en unknown
- 1974-06-21 NO NO742274A patent/NO742274L/no unknown
- 1974-06-22 ES ES427568A patent/ES427568A1/en not_active Expired
- 1974-06-24 HU HUAI236A patent/HU168582B/hu unknown
- 1974-06-24 DK DK338574A patent/DK338574A/da unknown
- 1974-06-24 BE BE145822A patent/BE816786R/en active
- 1974-06-24 FI FI1930/74A patent/FI193074A/fi unknown
- 1974-06-24 DD DD179409A patent/DD112519A5/xx unknown
- 1974-06-25 JP JP49072710A patent/JPS5049026A/ja active Pending
- 1974-06-25 FR FR7422137A patent/FR2234558B2/fr not_active Expired
- 1974-06-25 NL NL7408556A patent/NL7408556A/xx unknown
- 1974-06-25 BR BR5213/74A patent/BR7405213D0/en unknown
- 1974-06-25 AU AU70491/74A patent/AU7049174A/en not_active Expired
Also Published As
| Publication number | Publication date |
|---|---|
| NL7408556A (en) | 1974-12-30 |
| FI193074A (en) | 1974-12-26 |
| DK338574A (en) | 1975-02-24 |
| DD112519A5 (en) | 1975-04-12 |
| FR2234558A2 (en) | 1975-01-17 |
| BE816786R (en) | 1974-12-24 |
| SE7407899L (en) | 1974-12-27 |
| AU7049174A (en) | 1976-01-08 |
| DE2429647A1 (en) | 1975-01-16 |
| HU168582B (en) | 1976-06-28 |
| JPS5049026A (en) | 1975-05-01 |
| FR2234558B2 (en) | 1977-02-11 |
| BR7405213D0 (en) | 1975-01-28 |
| ES427568A1 (en) | 1976-07-16 |
| CA1037366A (en) | 1978-08-29 |
| ZA743995B (en) | 1975-06-25 |
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