DE2019778A1 - Detmn of ribonucleic acid distribution in cells - Google Patents
Detmn of ribonucleic acid distribution in cellsInfo
- Publication number
- DE2019778A1 DE2019778A1 DE19702019778 DE2019778A DE2019778A1 DE 2019778 A1 DE2019778 A1 DE 2019778A1 DE 19702019778 DE19702019778 DE 19702019778 DE 2019778 A DE2019778 A DE 2019778A DE 2019778 A1 DE2019778 A1 DE 2019778A1
- Authority
- DE
- Germany
- Prior art keywords
- ribonucleic acid
- ribonucleic acids
- solution
- thionine
- detmn
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 229920002477 rna polymer Polymers 0.000 title claims abstract description 16
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 30
- 102000007999 Nuclear Proteins Human genes 0.000 claims abstract description 5
- 108010089610 Nuclear Proteins Proteins 0.000 claims abstract description 5
- KUUVQVSHGLHAKZ-UHFFFAOYSA-N thionine Chemical compound C=1C=CC=CSC=CC=1 KUUVQVSHGLHAKZ-UHFFFAOYSA-N 0.000 claims abstract description 5
- 229950003937 tolonium Drugs 0.000 claims abstract description 5
- 230000001086 cytosolic effect Effects 0.000 claims abstract description 3
- 230000003834 intracellular effect Effects 0.000 claims abstract description 3
- 238000000034 method Methods 0.000 claims description 21
- 239000000243 solution Substances 0.000 claims description 11
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- 241001465754 Metazoa Species 0.000 claims description 6
- 239000000981 basic dye Substances 0.000 claims description 4
- 239000007853 buffer solution Substances 0.000 claims description 4
- 230000001476 alcoholic effect Effects 0.000 abstract 2
- RNVCVTLRINQCPJ-UHFFFAOYSA-N o-toluidine Chemical compound CC1=CC=CC=C1N RNVCVTLRINQCPJ-UHFFFAOYSA-N 0.000 abstract 2
- 230000002759 chromosomal effect Effects 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 10
- 238000002360 preparation method Methods 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 241000196324 Embryophyta Species 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 230000003179 granulation Effects 0.000 description 4
- 238000005469 granulation Methods 0.000 description 4
- 230000003458 metachromatic effect Effects 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- 210000000349 chromosome Anatomy 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 239000000975 dye Substances 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Natural products CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- 241000256128 Chironomus <genus> Species 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- 229960000583 acetic acid Drugs 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- WYICGPHECJFCBA-UHFFFAOYSA-N dioxouranium(2+) Chemical compound O=[U+2]=O WYICGPHECJFCBA-UHFFFAOYSA-N 0.000 description 2
- 239000000834 fixative Substances 0.000 description 2
- 229960004279 formaldehyde Drugs 0.000 description 2
- 235000019256 formaldehyde Nutrition 0.000 description 2
- 230000011278 mitosis Effects 0.000 description 2
- KMUONIBRACKNSN-UHFFFAOYSA-N potassium dichromate Chemical compound [K+].[K+].[O-][Cr](=O)(=O)O[Cr]([O-])(=O)=O KMUONIBRACKNSN-UHFFFAOYSA-N 0.000 description 2
- 244000178870 Lavandula angustifolia Species 0.000 description 1
- 235000010663 Lavandula angustifolia Nutrition 0.000 description 1
- 241001599018 Melanogaster Species 0.000 description 1
- UEZVMMHDMIWARA-UHFFFAOYSA-N Metaphosphoric acid Chemical compound OP(=O)=O UEZVMMHDMIWARA-UHFFFAOYSA-N 0.000 description 1
- 230000006819 RNA synthesis Effects 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- YOWZJZJLXUQHGF-UHFFFAOYSA-M [7-(dimethylamino)phenothiazin-3-ylidene]-dimethylazanium;dimethyl-[7-(methylamino)phenothiazin-3-ylidene]azanium;dichloride Chemical compound [Cl-].[Cl-].C1=CC(=[N+](C)C)C=C2SC3=CC(NC)=CC=C3N=C21.C1=CC(=[N+](C)C)C=C2SC3=CC(N(C)C)=CC=C3N=C21 YOWZJZJLXUQHGF-UHFFFAOYSA-M 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- KRVSOGSZCMJSLX-UHFFFAOYSA-L chromic acid Substances O[Cr](O)(=O)=O KRVSOGSZCMJSLX-UHFFFAOYSA-L 0.000 description 1
- UFMZWBIQTDUYBN-UHFFFAOYSA-N cobalt dinitrate Chemical compound [Co+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O UFMZWBIQTDUYBN-UHFFFAOYSA-N 0.000 description 1
- 229910001981 cobalt nitrate Inorganic materials 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- AWJWCTOOIBYHON-UHFFFAOYSA-N furo[3,4-b]pyrazine-5,7-dione Chemical compound C1=CN=C2C(=O)OC(=O)C2=N1 AWJWCTOOIBYHON-UHFFFAOYSA-N 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 1
- 239000001102 lavandula vera Substances 0.000 description 1
- 235000018219 lavender Nutrition 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000006140 methanolysis reaction Methods 0.000 description 1
- 230000011987 methylation Effects 0.000 description 1
- 238000007069 methylation reaction Methods 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 210000003079 salivary gland Anatomy 0.000 description 1
- 229920003002 synthetic resin Polymers 0.000 description 1
- 239000000057 synthetic resin Substances 0.000 description 1
- HNONEKILPDHFOL-UHFFFAOYSA-M tolonium chloride Chemical compound [Cl-].C1=C(C)C(N)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 HNONEKILPDHFOL-UHFFFAOYSA-M 0.000 description 1
- 229910002007 uranyl nitrate Inorganic materials 0.000 description 1
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
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5094—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for blood cell populations
-
- 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
- G01N2001/307—Fixative compositions non-toxic, no Hg, no formaldehyde
Abstract
Description
Verfahren zur Bestimmung der Zellverteilunz von Ribonukleinsäuren Die vorliegende Erfindung bezieht sich auf ein metachromatisches Verfahren zur Bestimmung der intrazellulären Verteilung der Ribonukleinsäuren in Tier- und Pflanzengeweben. Method for determining the cell distribution of ribonucleic acids The present invention relates to a metachromatic method of determination the intracellular distribution of ribonucleic acids in animal and plant tissues.
Die Kenntnis der Ribonukleinsäureverteilung in den Zellen, und die bei dieser Verteilung vorkommenden Veränderungen, sowohl bei der Mitose als auch bei verschiedenen tathologischen Prozesses, hat eine hervorragende Bedeutsamkeit für die Zellbioiogie.Knowledge of the ribonucleic acid distribution in cells, and the changes occurring in this distribution, both in mitosis and in various process of tathology, has a great significance for cell biology.
Zur Bestimmung der Ribonukleinsäureverteilung sind bereib zahlreiche Verfahren bekannt. Es gibt, z. B.There are numerous available for determining the ribonucleic acid distribution Procedure known. There are e.g. B.
metachtomatische Methoden, welche die Eigenschaft des Toluidinblaus und des Azurs I und II benutzen, rotfarbige Verbindungen mit der Ribonukleinsäure und Fettstoffen zu bilden; diese Methoden sind aber unspezifisch und unselektiv. Man benutzt noch zum selben Zwecke weiterhin die Eigenschaften des Pyronins, die riboneukleinischen Zellstrukturen rot zu färben; diese Methode gestattet jedoch den Nachweis der chromosomischen Ribonukleinsäure nicht und gibt keine Auskunft über die Nukleolstruktur.metachomatic methods showing the property of toluidine blue and Azure I and II use red-colored compounds with ribonucleic acid and fatty substances to build; however, these methods are unspecific and unselective. The properties of the are still used for the same purpose Pyronins to stain ribonuclein cell structures red; this method allowed however, the detection of the chromosomic ribonucleic acid does not provide any information about the nucleolar structure.
Ebenfalls kennt man noch elektromikroskopische und autoradiographische Methoden, bei welchen man das tritiumhaltige tridin (H3) als Vorgänger in der Ribonukleinsäurebiosynthese benutzt, beide Verfahren setzen jedoch eine teuere Apparatur voraus und sind schwierig-durchzufiüiren.Electromicroscopic and autoradiographic ones are also known Methods in which the tritiated tridine (H3) is used as a precursor in ribonucleic acid biosynthesis are used, but both methods require expensive equipment and are difficult to carry out.
Die vorliegende Erfindung beseitigt die Nachteile der erwähnten Methoden dadurch,.dass die Schnitte von fixierten Tier-und Pflanzengeweben mit basischen Farbstoffen aus der Thioningruppe in hydroalkoholischer 0,005 bis 0,01 gew.°%iger Pufferlösung gefärbt, mit Methanol, welches~0,8 Vol.% Salzsäure enthält und nachher mit einer hydroalkoholischen 0,5 bis 1gew.0/ igen Essiguranyllösung behandelt werden.The present invention overcomes the disadvantages of the aforementioned methods thereby, that the sections of fixed animal and plant tissues with basic Dyes from the thionine group in hydroalcoholic 0.005 to 0.01% by weight Buffer solution colored, with methanol, which contains ~ 0.8% by volume of hydrochloric acid and afterwards be treated with a hydroalcoholic 0.5 to 1% by weight aceticuranyl solution.
Das Verfahren stützt sich auf die Eigenschaft der basischen Farbstoffe aus der Ehioningruppe und besonders des 0-Toluidinblaus, alle ribonukleinischen ZellstruXturen auf vorher mit hydrochlorischem Methanol behandelten miktoskopischen Präparaten rot zu färben, wonach eine Uranylionenbehandiung'durchgeführt wird.The process is based on the property of basic dyes from the ehionin group and especially the 0-toluidine blue, all ribonucleinic Cell structures on microscopic microscopes previously treated with hydrochloric methanol To color preparations red, after which a uranyl ion treatment is carried out.
Die Spiazifität -des Verfahrens besteht darin, dass durch Behandlung -der Tier- -oder Pflanzengewebeschnitte mit hydrochlorischem methanol die Gruppierungen -COOH durch Methylierung) und -0-SODH (durch Methan-olyse) aus den sauren Poly-~ sacchariden blockiert werden, wodurch verhindert wird, dass diese an metachromatischen Reaktionen teilnehmen.The practicality of the procedure is that through treatment -the animal or plant tissue sections with hydrochloric methanol the groupings -COOH by methylation) and -0-SODH (by methanolysis) from the acidic poly- ~ saccharides are blocked, which prevents them from becoming metachromatic Reactions to participate.
Durch die Anwendung des Farbstoffs in hydroalkoholischer Lösung wi-rd weiterbin die Möglichkeit beseitigt, dass die sauren Zellfettstoffe metachromatisch -reagieren, da auch die geringsten Alkoholmengen die vondiesen Fettstoffen gebildete Metachromasie vernichten. Durch Verwendung der Uranylionenpräparate wird bei den Arbeitsbedingungen dieses Verfahrens gewährleistet, dass nur die Zellribonukleinsäure- wegen der in ihrem Molekä: enthaltenen Phosphatgruppierungen - - beständige metachromatische Komplexe, d. h. rotfarbige Komplexe, mit dem Farbstoff -bildet, während die anderen nicht ribonukleischen-Zellstrukture#sich orthochromatisch, d. d. h. hellblau, färben.By using the dye in a hydroalcoholic solution wi-rd Furthermore, the possibility is eliminated that the acidic cell fat substances are metachromatic -react, since even the smallest amounts of alcohol are those formed by these fatty substances Destroy metachromasia. By using the uranyl ion preparations, the The working conditions of this process ensure that only the cell ribonucleic acid because of the phosphate groups contained in their molecule - - permanent metachromatic Complexes, d. H. red-colored complexes, with the dye -forms, while the other non-ribonuclear cell structure # is orthochromatic, d. d. H. light blue, color.
Zum Testen des erfindungsgemässen Verfahrens wurden Mäuse-und Rattenleber, Rattenhepatom, Larven von Chironomus tetans und Drosophilla melanogaster und Wurzelspitzen verschiedener P-flanzenarten verwendet. -Weitere Vorteile und Merkmale der Erfindung ergeben sich aus, der folgenden Beschreibung anhand eines Ausführungsbeispiels: Kleine Teilchen von Tiergeweben (Volumen 3 1 - 2 mm3), Larven oder Wurzeispitzen (2 - 3 mm lang) werden 12 bis 24 Stunden bei 5 bis 8° C mit einem der unten angegebenen Fixiermittel behandelt: I. 0,5gew.%ige Chromsäurelösung 8 ml Eisessig@ 0,4 ml 4O%iges Formol 1,6 ml II. Kobaltnitrat 0,1 g Uranylnitrat 0,1 g 5°%ige Kaliumbichromatlösung 3 ml Äthanol 6 ml Ä-thyläther 2 ml 4O%iges Formol 4 ml - Eisessig 1 ml.To test the method according to the invention, mouse and rat liver, Rat hepatoma, larvae of Chironomus tetans and Drosophilla melanogaster and root tips different types of plants are used. -Other advantages and features of the invention result from the following description based on an exemplary embodiment: Small Particles of animal tissue (volume 3 1 - 2 mm3), larvae or root tips (2 - 3 mm long) are 12 to 24 hours at 5 to 8 ° C with one of the below Fixative treated: I. 0.5% by weight chromic acid solution 8 ml glacial acetic acid @ 0.4 ml 40% Formol 1.6 ml II. Cobalt nitrate 0.1 g uranyl nitrate 0.1 g 5% potassium dichromate solution 3 ml ethanol 6 ml ethyl ether 2 ml 4O% formol 4 ml - glacial acetic acid 1 ml.
Das - Fixiermittel wird bei der Benutzung hergestellt, unter Anwendung von analysenreinen Substanzen.The fixative is made in use, under application of analytically pure substances.
Nach Fixierung werden die Teile mit kaltem Wasser gewaschen, entwässert, in Paraffin eingelagert und gemäss der üblichen Technik Schnitte von 3 bis 5 Mikron hergestellt. Nach Entparaffinieren werden die Präparate in zwei Gruppen A und B geteilt.After fixing, the parts are washed with cold water, drained, stored in paraffin and sections of 3 to 5 microns according to the usual technique manufactured. After dewaxing, the preparations are divided into two groups, A and B. divided.
Die Präparate der Gruppe A werden 20 Minuten lang bei 600 a, in 0,1 N HCl-Lösung gebracht, gewaschen und entwässert.The preparations of group A are for 20 minutes at 600 a, in Brought 0.1 N HCl solution, washed and drained.
Sowohl die auf diese Weise behandelten Präparate der Gruppe A, wie auch diejenigen der Gruppe B werden der selben Behandlung unterworfen: Die Präparate werden in drei aufeinanderfolgende Bäder von absolutem Methanol gebracht und anschliessend 1 Stunde bei 37° C in absolutem Methanol, welcher 0,8 ml konz.Both the preparations of group A treated in this way, as well as those in group B are also subjected to the same treatment: the preparations are placed in three successive baths of absolute methanol and then 1 hour at 37 ° C in absolute methanol, which 0.8 ml of conc.
HCl (D = 1,19) in100 ml Alkohol enthält, stehengelassen. Nach dieser Zeitspanne werden die Präparate herausgenommen, wenigstens 10 Mal mit etwas gekühltem destilliertem Wasser bei 5 bis 80 C gewaschen und nachher in eine 0,5 bis 1°ige Essiguranyllösung in 30%igem Methanol eingetragen und 1 Stunde bei Zimmertemperatur stehengelassen; darauf werden die Schnitte wenigstens 10 Mal mit gekühltem Wasser bei 5 bis 80 C gewaschen und 10 Minuten bei Zimmertemperatur in einer'Pufferlösung vom pH-Wert 3,4 bis 3,6 stehengelassen1 die wie folgt hergestellt wird: 0,1 fl krist. Essignatriumlösung 12,5 ml 0,1 M Phosphorsäurelösung 8 ml 0,1 X Zitronensäurelösung 4 ml Bidestilliertes Wasser 45,5 ml Absolutes Methanol 30 ml (Der: pH-Wert wird bei der Lösung ohne Methanol eingestellt.) Schliesslich trägt man die Präparate direkt in die frisch hergestellte Farbstofflösung ein, welche durch Auflösen von 0,005 bis 0,01 gew.oigem 0-Toluidinblau (Merck) in obige Pufferlösung hergestellt wurde, und lässt 45 bis 48 Stunden bei Zimmertemperatur stehen. Dann werden die Präparate herausgeholt, in Wasser gewaschen und entwässert und nachher in ECaz74-dabalsam oder, vorzugsweise, in ein Kunstharz montiert.HCl (D = 1.19) in 100 ml of alcohol, left to stand. After this During this period the preparations are taken out, at least 10 times with something chilled Washed distilled water at 5 to 80 C and then in a 0.5 to 1 ° ige Essiguranyllösung registered in 30% methanol and 1 hour at room temperature ditched; then the cuts are made at least 10 times with chilled water Washed at 5 to 80 C and 10 minutes at room temperature in a buffer solution from pH 3.4 to 3.6 left to stand1, which is prepared as follows: 0.1 fl crystall. Acetate sodium solution 12.5 ml 0.1 M phosphoric acid solution 8 ml 0.1 X citric acid solution 4 ml bidistilled water 45.5 ml absolute methanol 30 ml (The: pH value becomes set in the solution without methanol.) Finally, the specimens are worn directly into the freshly made Dye solution, which through Dissolve 0.005 to 0.01% by weight of 0-toluidine blue (Merck) in the above buffer solution and leave to stand at room temperature for 45 to 48 hours. then the specimens are taken out, washed in water and dehydrated and afterwards in ECaz74-dabalsam or, preferably, mounted in a synthetic resin.
Die mikroskopische Untersuchung der auf diese Weise erhaltenen Präparate zeigt, dass alle ribonukleischen Zellstrukturen rot gefärbt sind. So erscheint die cytoplasmatische Ribonukleinsäure in Form von zahlreichen und winzigen rotgefärbten Granulationen. Die chromosomische Ribonukleinsäure ist gekennzelchnet durch rote Granulationen, die längs der lavendelblau gefärbten Chromosome angeordnet sind. Die Ribonukleinsäure aus den Riesenchromosomen der Speicheldrüsen von Chironomus und Drosophilla haben die Form von roten Granulationen, die in schrägen Streifen angeordnet sind. Die nukleoläre Ribonukleinsäure erscheint als rote Granulationen verschiedener Grössen, die in die nukleoläre Matrix - welche auch rot gefärbt ist -eingelagert sind.Microscopic examination of the preparations obtained in this way shows that all ribonuclear cell structures are stained red. This is how it appears cytoplasmic ribonucleic acid in the form of numerous and tiny red-colored Granulations. The chromosomic ribonucleic acid is marked with red Granulations arranged along the lavender blue colored chromosomes. The ribonucleic acid from the giant chromosomes of the salivary glands of Chironomus and Drosophilla are in the form of red granulations that appear in oblique stripes are arranged. The nucleolar ribonucleic acid appears as red granulations different sizes, which are in the nucleolar matrix - which is also colored red -are stored.
Das Verfahren kann vorteilhaft zum Studium des Nukleolverhaltens während der Mitose angewandt werden, was ein aktuelles Problem in der Zellbiologie darstellt; es kann wichtige Aufschlüsse über die Form-, Volumen-, Struktur- und Anzahlvariation der Nukleole in den krebserzeugenden Prozessen geben, und kann präzisere Ergebnisse bei der Funktionsweise der nakleol-organisierenden Chromosome, wie auch über die Herkubft der ikronukleole, Nukleoline und Karyosome erbringen.The method can be beneficial for studying nucleolic behavior during mitosis, which is a current problem in cell biology; it can provide important information about the variation in shape, volume, structure and number of the nucleoli in the carcinogenic processes, and can be more precise Results in the functioning of the nakleol-organizing chromosomes, as well via the origins of the micronucleols, nucleolins and karyosomes.
In Betracht gezogene Druckschriften: L. Lison: Ristochimie et Cytochimie animales. Principes et methodes - Gauthier-Villars, Paris, 1960, Vol. I, pag. 280.- 300 A.G. Everson Pearse: Ristochemistry theoretical and applied J. si A. Churchill London, 1961, pag. 248 - 260 J.A. Bergeron si M. Singer: Metachromasy; S. Biophysic and Biochem. Cytol,, 1958, 4, pag. 433-45?.References considered: L. Lison: Ristochimie et Cytochimie animales. Principes et methodes - Gauthier-Villars, Paris, 1960, Vol. I, p. 280.- 300 A.G. Everson Pearse: Ristochemistry theoretical and applied J. si A. Churchill London, 1961, p. 248-260 J.A. Bergeron and M. Singer: Metachromasy; S. Biophysic and Biochem. Cytol, 1958, 4, p. 433-45 ?.
Claims (3)
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19702019778 DE2019778B2 (en) | 1970-04-23 | 1970-04-23 | METHOD FOR DETERMINING THE INTRACELLULAR DISTRIBUTION OF RIBONUCLEIC ACIDS |
| FR7018020A FR2088100A1 (en) | 1970-04-23 | 1970-05-19 | Detmn of ribonucleic acid distribution in cells |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19702019778 DE2019778B2 (en) | 1970-04-23 | 1970-04-23 | METHOD FOR DETERMINING THE INTRACELLULAR DISTRIBUTION OF RIBONUCLEIC ACIDS |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| DE2019778A1 true DE2019778A1 (en) | 1971-11-18 |
| DE2019778B2 DE2019778B2 (en) | 1973-02-08 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DE19702019778 Granted DE2019778B2 (en) | 1970-04-23 | 1970-04-23 | METHOD FOR DETERMINING THE INTRACELLULAR DISTRIBUTION OF RIBONUCLEIC ACIDS |
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| Country | Link |
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| DE (1) | DE2019778B2 (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2478317A1 (en) * | 1980-03-12 | 1981-09-18 | Kass Lawrence | METHOD AND COMPOSITION FOR DETERMINING INDIVIDUAL CATEGORIES OF LEUKOCYTES BY DIFFERENTIAL SORPTION OF AT LEAST ONE METACHROMATIC DYE |
| US5134662A (en) * | 1985-11-04 | 1992-07-28 | Cell Analysis Systems, Inc. | Dual color camera microscope and methodology for cell staining and analysis |
| US5202931A (en) * | 1987-10-06 | 1993-04-13 | Cell Analysis Systems, Inc. | Methods and apparatus for the quantitation of nuclear protein |
| US5281517A (en) * | 1985-11-04 | 1994-01-25 | Cell Analysis Systems, Inc. | Methods for immunoploidy analysis |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5109429A (en) * | 1985-11-04 | 1992-04-28 | Cell Analysis Systems,Inc. | Apparatus and method for analyses of biological specimens |
-
1970
- 1970-04-23 DE DE19702019778 patent/DE2019778B2/en active Granted
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2478317A1 (en) * | 1980-03-12 | 1981-09-18 | Kass Lawrence | METHOD AND COMPOSITION FOR DETERMINING INDIVIDUAL CATEGORIES OF LEUKOCYTES BY DIFFERENTIAL SORPTION OF AT LEAST ONE METACHROMATIC DYE |
| US5134662A (en) * | 1985-11-04 | 1992-07-28 | Cell Analysis Systems, Inc. | Dual color camera microscope and methodology for cell staining and analysis |
| US5281517A (en) * | 1985-11-04 | 1994-01-25 | Cell Analysis Systems, Inc. | Methods for immunoploidy analysis |
| US5202931A (en) * | 1987-10-06 | 1993-04-13 | Cell Analysis Systems, Inc. | Methods and apparatus for the quantitation of nuclear protein |
Also Published As
| Publication number | Publication date |
|---|---|
| DE2019778B2 (en) | 1973-02-08 |
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