Automatische Silberkornzählung in der Einzelzell-Autoradiographie Eine neue photometrische Methode für die quantitative Autoradiographie

Dormer, Peter G.; Brinkmann, W; Stieber, Anna; Stich, W.

doi:10.1007/bf01727572

Cited by 35 publications

(4 citation statements)

References 7 publications

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“…Only after subtraction of this 'machine background' is the linearity of grain count with photometer reading observed. Similar plots have been presented byDormer et al (1966),Dormer (1967), while Goldstein & Williams (1971) have also confirmed this relationship. Using AR-10 stripping film,Dormer (1967) has shown that the linear relationship holds up to densities of seventy developed grains per 10 pm2.…”

supporting

confidence: 81%

Photometrie measurements of grain density in autoradiographs

Rogers

1972

Journal of Microscopy

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SUMMARY A technique is described for measuring the grain densities in areas of emulsion from 10 to 200 μm2, using vertical incident illumination which converts the silver grains into bright specks on a black background. The light reflected by the silver grains in the measuring area is integrated with a photometer to give a reading that is directly proportional to the number of grains present over a wide range of densities. Speeds of 200 counts/h can be maintained on suitable material throughout a working day without loss of accuracy. The characteristics of the method, the selection of equipment, and some sources of artefacts are discussed.

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confidence: 81%

Photometrie measurements of grain density in autoradiographs

Rogers

1972

Journal of Microscopy

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“…The density of silver grains was determined by leading a measuring diaphragm of 552 µ2 around the central and the periportal areas of the liver THE JOURNAL OF CELL BIOLOGY • VOLUME 47, 1970 • pages 285-289 lobule and measuring the light reflected by the silver grains in a microscope equipped with a photomultiplier (4) . The pulses of the photomultiplier were recorded as "working units" by a Knott analog digital counter .…”

Section: Methodsmentioning

confidence: 99%

The Distribution of Albumin Synthesis Throughout the Liver Lobule

Schreiber

Lesch

Weinssen

et al. 1970

The Journal of Cell Biology

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INTRODUCTIONThe membrane-attached polysomes of the liver cells are assumed to be the intracellular site of albumin synthesis (for recent critical review see Campbell (1)) . The histological localization of albumin by fluorescent antisera has been described repeatedly (3, 5-7) . It is not possible, however, to decide whether albumin localized in the cell by this technique is newly synthesized, or taken up by the cells from the blood, or simply adsorbed during preparation .In radioautographs we observed predominant labeling of the periportal area of the liver lobules after intraportal injection of leucine-3H, whereas intracaval injection led to uniform labeling throughout the lobule . The way of administration of the tracer thus permits a study of protein synthesis of selected regions of the liver lobule .Recently, the isolation of radiochemically pure albumin from liver homogenates and a method to determine the proportion of albumin to total protein synthesis have been described (9) .In this report the ratio of albumin to total protein synthesis after intraportal injection was compared to that after intracaval injection . By combining these results with the radioautographic observations, it was possible to decide whether or not albumin synthesis was distributed uniformly throughout the liver lobule . MATERIALS AND METHODSAnimals and biochemical methods have been described previously (8, 9) .For radioautography small pieces of liver were fixed in 0.75 M sodium phosphate buffer of pH 7 .0 with 4% formaldehyde for 24 hr at 4°C, dehydrated in ethanol, and embedded in paraffin blocks . Slices of 5 µ and control slices from unlabeled livers were cut and transferred to the same microscopical slides . Paraffin was removed by alcohol, methylbenzoate, and xylene. The slices were washed twice for 20 min in 0 .76 mm nonradioactive leucine and then dipped into Ilford K5 liquid emulsion (Ilford Ltd ., Ilford, Essex, England) . After 24 days' exposure at 18°C and after photographic development, radioautographs were stained through the emulsion with Mayer's hemalum, made transparent by 70% ethanol with 4 vol % of 1 N HCI, rinsed in running tap water, and dried .The density of silver grains was determined by leading a measuring diaphragm of 552 µ2 around the central and the periportal areas of the liver THE JOURNAL OF CELL BIOLOGY • VOLUME 47, 1970 • pages 285-289 285 on May 13, 2018 jcb.rupress.org Downloaded from http://doi.org/10.1083/jcb.47.1.285 Published Online: 1 October, 1970 | Supp Info:lobule and measuring the light reflected by the silver grains in a microscope equipped with a photomultiplier (4) . The pulses of the photomultiplier were recorded as "working units" by a Knott analog digital counter . Two animals were measured after intraportal, and two after intracaval injection of the label . Almost identical values were obtained within each group . The obtained values were averaged after subtraction of the background, which was determined with unlabeled slices . After intraportal injection, 100 positions of the diaph...

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“…All cell smears were proved with a computerized scanning microspectrophotometer based on a Univar photomicroscope (Reichert Optische Werke Vienna) using the 'Autoradiography' computer program package according to Dormer (1967), Dormer & Brinkmann (1968) and Dormer et al (1966, using an interactive computer controlled calibration curve which, in incident light illumination with a blue interference filter (wavelength 435 nm), independently provides the relationship between the silver grain density and the photometer reading. Microscale-blocks (Autoradiographic 3H-microscales, Amersham International plc, Code RA 501) served as standards to produce the calibration curve.…”

Section: Quantitative Evaluation Of the Autoradiogramsmentioning

confidence: 99%

Demonstration of pseudorabies virus DNA in the mouse inner ear by an in situ nucleic acid hybridization technique in plastic embedded bony material

Falser

Bandtlow

Haus³

et al. 1986

Journal of Microscopy

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SUMMARYThis investigation is concerned with the possibility of identifying viral DNA using the in situ DNA hybridization method in methylmethacrylate-embedded material. As an experimental model we chose viral labyrinthitis produced by intranasal infection of the mouse with pseudorabies virus. Fixation and embedding methods specially adapted to this procedure and bony histology preparation technique (specimens by grinding or micromilling) made it possible to identify viral DNA directly morphologically and virologically in the inner ear. Quantitative microphotometric analyses of trans-sagittal sections of the entire skull after in situ DNA hybridization are presented and discussed here as an explicit method of investigating the path of distribution of viral DNA in the brain and the inner ear.

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Automatische Silberkornzählung in der Einzelzell-Autoradiographie Eine neue photometrische Methode für die quantitative Autoradiographie

Cited by 35 publications

References 7 publications

Photometrie measurements of grain density in autoradiographs

Photometrie measurements of grain density in autoradiographs

The Distribution of Albumin Synthesis Throughout the Liver Lobule

Demonstration of pseudorabies virus DNA in the mouse inner ear by an in situ nucleic acid hybridization technique in plastic embedded bony material

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