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The Combined Use of Autoradiographic and Electron Microscopic Techniques for Studies on Ultra-Thin Sections of Tritium-Labelled Cells of the Intestinal Epithelium; Emploi Combine de l'Autoradiographie et du Microscope Electronique pour L'Etude de Coupes Ultra-Fines de Cellules Tritiees de l'Epithelium Intestinal; 0421 043e 0432 043c 0435 0414 ; Empleo Combinado de Tecnicas de Autorradiografia y de MicroscopiA Electronica para Estudiar Cortes Ultrafinos de Celulas Tritiadas del Epitelio Intestinal

Abstract

The high resolution now obtainable in sectioned cells studied in the electron microscope offers some promise of visualizing some of the intracellular events leading to the synthesis of proteins. The specificity of incorporation of tritiated thymidine into DNA provides a system which can be accurately followed with respect to time and localization within the cell. The work to be reported here demonstrates that it is possible to study ultra-thin sections of labelled cells in the electron microscope, to remove the specimen after suitable electron micrographs have been taken, to apply a coating of emulsion sufficiently thick to obtain an autoradiograph but still thin enough' to permit re-examination of the specimen in the electron microscope and to obtain electron micrographic images of autoradiography of cells previously recorded. The autoradiographic emulsion was applied as follows: a thin film was formed by dipping a small wire loop into liquid emulsion and transferring the film by passing the loop over the mounted specimen which was fixed to the top of a small plastic peg. Adequate control can be achieved by taking a one-half micron section after each ultra-thin section during sectioning, mounting it on a glass slide and applying conventional autoradiographic techniques. Because of  More>>
Authors:
Hampton, J. C.; [1]  Quastler, H. [2] 
  1. Division of Experimental Biology, Baylor University College of Medicine, TX (United States)
  2. Brookhaven National Laboratory, Upton, Long Island, NY (United States)
Publication Date:
Feb 15, 1962
Product Type:
Conference
Resource Relation:
Conference: Symposium on the Detection and Use of Tritium in the Physical and Biological Sciences, Vienna (Austria), 3-10 May 1961; Other Information: 5 refs., 6 figs.; Related Information: In: Tritium in the Physical and Biological Sciences. Vol. II. Proceedings of the Symposium on the Detection and Use of Tritium in the Physical and Biological Sciences| 456 p.
Subject:
38 RADIATION CHEMISTRY, RADIOCHEMISTRY AND NUCLEAR CHEMISTRY; 60 APPLIED LIFE SCIENCES; 63 RADIATION, THERMAL, AND OTHER ENVIRONMENTAL POLLUTANT EFFECTS ON LIVING ORGANISMS AND BIOLOGICAL MATERIALS; AUTORADIOGRAPHY; DNA; ELECTRON MICROSCOPES; ELECTRONS; EMULSIONS; EPITHELIUM; IMAGES; LABELLED COMPOUNDS; PLASTICS; PROTEINS; THIN FILMS; THYMIDINE; TRITIUM
OSTI ID:
22188133
Research Organizations:
International Atomic Energy Agency, Vienna (Austria); Joint Commission on Applied Radioactivity of the International Council of Scientific Unions, Paris (France)
Country of Origin:
IAEA
Language:
English
Other Identifying Numbers:
Other: ISSN 0074-1884; TRN: XA13M3692012634
Submitting Site:
INIS
Size:
page(s) 103-110
Announcement Date:
Feb 06, 2014

Citation Formats

Hampton, J. C., and Quastler, H. The Combined Use of Autoradiographic and Electron Microscopic Techniques for Studies on Ultra-Thin Sections of Tritium-Labelled Cells of the Intestinal Epithelium; Emploi Combine de l'Autoradiographie et du Microscope Electronique pour L'Etude de Coupes Ultra-Fines de Cellules Tritiees de l'Epithelium Intestinal; 0421 043e 0432 043c 0435 0414 ; Empleo Combinado de Tecnicas de Autorradiografia y de MicroscopiA Electronica para Estudiar Cortes Ultrafinos de Celulas Tritiadas del Epitelio Intestinal. IAEA: N. p., 1962. Web.
Hampton, J. C., & Quastler, H. The Combined Use of Autoradiographic and Electron Microscopic Techniques for Studies on Ultra-Thin Sections of Tritium-Labelled Cells of the Intestinal Epithelium; Emploi Combine de l'Autoradiographie et du Microscope Electronique pour L'Etude de Coupes Ultra-Fines de Cellules Tritiees de l'Epithelium Intestinal; 0421 043e 0432 043c 0435 0414 ; Empleo Combinado de Tecnicas de Autorradiografia y de MicroscopiA Electronica para Estudiar Cortes Ultrafinos de Celulas Tritiadas del Epitelio Intestinal. IAEA.
Hampton, J. C., and Quastler, H. 1962. "The Combined Use of Autoradiographic and Electron Microscopic Techniques for Studies on Ultra-Thin Sections of Tritium-Labelled Cells of the Intestinal Epithelium; Emploi Combine de l'Autoradiographie et du Microscope Electronique pour L'Etude de Coupes Ultra-Fines de Cellules Tritiees de l'Epithelium Intestinal; 0421 043e 0432 043c 0435 0414 ; Empleo Combinado de Tecnicas de Autorradiografia y de MicroscopiA Electronica para Estudiar Cortes Ultrafinos de Celulas Tritiadas del Epitelio Intestinal." IAEA.
@misc{etde_22188133,
title = {The Combined Use of Autoradiographic and Electron Microscopic Techniques for Studies on Ultra-Thin Sections of Tritium-Labelled Cells of the Intestinal Epithelium; Emploi Combine de l'Autoradiographie et du Microscope Electronique pour L'Etude de Coupes Ultra-Fines de Cellules Tritiees de l'Epithelium Intestinal; 0421 043e 0432 043c 0435 0414 ; Empleo Combinado de Tecnicas de Autorradiografia y de MicroscopiA Electronica para Estudiar Cortes Ultrafinos de Celulas Tritiadas del Epitelio Intestinal}
author = {Hampton, J. C., and Quastler, H.}
abstractNote = {The high resolution now obtainable in sectioned cells studied in the electron microscope offers some promise of visualizing some of the intracellular events leading to the synthesis of proteins. The specificity of incorporation of tritiated thymidine into DNA provides a system which can be accurately followed with respect to time and localization within the cell. The work to be reported here demonstrates that it is possible to study ultra-thin sections of labelled cells in the electron microscope, to remove the specimen after suitable electron micrographs have been taken, to apply a coating of emulsion sufficiently thick to obtain an autoradiograph but still thin enough' to permit re-examination of the specimen in the electron microscope and to obtain electron micrographic images of autoradiography of cells previously recorded. The autoradiographic emulsion was applied as follows: a thin film was formed by dipping a small wire loop into liquid emulsion and transferring the film by passing the loop over the mounted specimen which was fixed to the top of a small plastic peg. Adequate control can be achieved by taking a one-half micron section after each ultra-thin section during sectioning, mounting it on a glass slide and applying conventional autoradiographic techniques. Because of the fragile nature of ultra-thin sections, the yield of successful autoradiographs is quite low. In spite of this objection it is expected that with improvements in skill and techniques the method will facilitate a better understanding of vital cell processes. (author) [French] Le haut degre de resolution que l'on peut maintenant obtenir dans l'etude des coupes de cellules au microscope electronique offre quelque possiblite d'observer certains des processus intracellulaires qui menent a la synthese des proteines. Le fait que la thymidine tritiee s'incorpore a l'ADN de facon specifique permet de suivre avec precision le deroulement d'un autre processus dans le temps et leur localisation dans la cellule. Les travaux presentes dans ce memoire demontrent qu'il est possible d'etudier au microscope electronique des coupes ultrafines de cellules marquees, de recuperer l'echantillon apres avoir pris des micrographies electroniques convenables, de l'enduire d'une emulsion suffisamment epaisse pour obtenir une autoradiographie, mais tout de meme assez mince pour permettre un nouvel examen au microscope electronique et obtenir des images micrographiques des autoradiographies deja faites. L'emulsion autoradiographique a ete appliquee comme suit: formation d'une mince pellicule sur une petite boucle de fil metallique plongee dans l'emulsion liquide, puis transfert de la pellicule par passage de la boucle sur l'echantillon monte, fixe a l'extremite d'une chevillette en plastique. On peut effectuer un controle satisfaisant en prenant, lorsqu'on fait les coupes, une coupe d'un demimicron apres chaque coupe ultrafine en la montant sur une lamelle de verre et en appliquant les techniques autoradiographiques classiques. En raison de la fragilite des coupes ultrafines, le nombre d'autoradiographies reussies est tres faible. Malgre cet inconvenient, on peut esperer qu'avec une plus grande experience et un perfectionnement des techniques, cette methode aidera a mieux comprendre les processus des cellules vitales. (author) [Spanish] Merced al elevado grado de resolucion que puede alcanzarse actualmente por observacion de cortes de celulas en el microscopio electronico, resulta posible estudiar algunos de los procesos intracelulares que conducen a la sintesis de las proteinas. La especificidad de la incorporacion de timidina tritiada en la molecula de acido desoxirribonucleico permite seguir detenidamente el desarrollo del proceso en funcion del tiempo y su localizacion dentro de la celula. Los trabajos descritos en la presente memoria demuestran que es posible estudiar por microscopia electronica cortes ultrafinos de celulas tritiadas, retirar la muestra una vez que se han tomado las micrografias electronicas adecuadas y aplicar una capa de emulsion suficientemente gruesa como para obtener una autorradiografia, pero, al mismo tiempo, bastante delgada como para permitir un nuevo examen de la muestra en el microscopio electronico y obtener micrografias electronicas de las autorradiografias de las celulas examinadas previamente. La emulsion autorradiografica se aplico en la siguiente forma: se formo primeramente una pelicula delgada sumergiendo un pequeno ojal de alambre en una emulsion liquida y trasladando la pelicula mediante el recurso de pasar el ojal sobre la muestra montado sobre un soporte, fijado a su vez en el extremo de una clavija de material plastico. El control adecuado se puede lograr tomando una seccion de media miera de espesor despues de cada operacion de corte, montandola sobre una placa de vidrio y aplicando luego las tecnicas autorradiograficas usuales. Debido a la fragilidad de los cortes ultrafinos, la proporcion de autorradiografias satisfactorias es bastante reducida. A pesar de este inconveniente, se espera que una vez perfeccionada la tecnica y adquirido experiencia en su manejo, este metodo permitira comprender mejor procesos que se verifican en las celulas vitales. (author) [Russian] Osvoennaja v nastojashhee vremja vozmozhnost' dostizhenija vysokoj razreshajushhej sposobnosti v izuchenii kletochnyh srezov pri pomoshhi jelektronnogo mikro- skopa otkryvaet puti k nabljudeniju za nekotorymi mezhkletochnymi javlenjami, vedushhimi k sintezu proteinov. Osobennosti pogloshhenija tritirovannogo timi- dina dezoksiribonukleinovoj kislotoj obrazujut sistemu, otkryvajushhuju vozmozhnost' tochnogo nabljudenija za vremenem ee proniknovenija i mestom ee sos- redotochenija v kletke. V dokladyvaemoj zdes' rabote pokazyvaetsja, chto sverh- tonkie srezy mechenyh kletochek mozhno izuchit' pri pomoshhi jelektronnogo mikroskopa, a zatem, posle poluchenija nuzhnoj mikrografii, vynut' obrazec, pokryt' ego plenkoj jemul'sii dostatochno tolstoj dlja poluchenija radioavtografii, no vse zhe dostatochno tonkoj dlja povtornogo izuchenija obrazca pod jelektronnym mikroskopom i dlja poluchenija novogo jelektronnogo mikrograficheskogo izo- brazhenija uzhe zasnjatogo ran'she radioavtografa kletok. Radioavtograficheskaja jemul'sija nakladyvalas' sledujushhim obrazom: pri pogruzhenii nebol'shoj provolochnoj petel'ki v zhidkuju jemul'siju na petle obrazovalas' tonkaja plenka, kotoraja perenosilas' jetoj petlej na prigotovlennyj obrazec, zakrepljavshijsja na okonechnosti nebol'shogo kolyshka iz plastmassy. Nadlezhashhij kontrol' mozhet proizvodit'sja putem snjatija sreza v polmikrona s kazhdogo sve- rhtonkogo sreza i nalozhenija ego na stekljannuju plastinku dlja semki obych- nym radioavtograficheskim metodom. Vvidu bol'shoj hrupkosti sverhtonkih srezov udaetsja poluchit' lish' nebol'shoe chislo udachnyh radioavtografov. Nesmotrja na jetot nedostatok, sleduet ozhidat', chto po mere uluchshenija tehniki i metoda jetot sposob oblegchit ponimanie vazhnyh zhiznennyh kletochnyh processov. (author)}
place = {IAEA}
year = {1962}
month = {Feb}
}