Auflichtphotometrie von Mikroautoradiogrammen für quantitative Einbaustudien an Einzelzellen

Dormer, Peter G.; Brinkmann, W

doi:10.1007/bf00546358

Cited by 14 publications

(2 citation statements)

References 9 publications

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“…In the present study, this was accomplished by sampling all relevant compartments within each cell and by normalization. For determination of absolute grain density, it is possible to include in each block a sample of polymerized labeled Araldite ( [8] see also Fig. 1); this serves as an internal standard, permitting correction of the photometric signal for random variations, e.g., thickness of section or emulsion and conditions of exposure or development,…”

Section: Quantitative Autoradiographymentioning

confidence: 99%

Secretory kinetics in the follicular cells of silkmoths during eggshell formation.

Blau¹,

Kafatos²

1978

The Journal of Cell Biology

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ABSTRAGTProcedures for quantitative autoradiography were used for studying the process of secretion of eggshell (chorion) proteins in the follicular epithelium of silkmoths. The method was based on photometric measurements of the reflectance of vertically illuminated autoradiographic silver grains. Results were analyzed and plotted by computer. Secretory kinetics were also determined by analysis of labeled proteins in physically separated epithelium and chorion. Rapid accumulation of radioactivity into "clumps" visualized by light microscope autoradiography and evidence from preliminary electron microscope autoradiography indicate that, within 2 min from the time of synthesis, labeled chorion proteins move to Golgi regions scattered throughout the cytoplasm. The proteins begin to accumulate in the apical area 10-20 rain later and to be discharged from the cell. The time for half-secretion is 20-25 min, and discharge is essentially complete 30-50 min after labeling. At the developmental stages examined, the kinetics of secretion appear to be similar for all proteins. Within the chorion the proteins rapidly assume a characteristic distribution, which varies for different developmental stages. Two relatively slow steps have been identified in secretion, associated with residence in Golgi regions and in the cell apex, respectively. By contrast, translocation of proteins across the cell and deposition of discharged proteins in the chorion are rapid steps.KEY WORDS secretion 9 quantitative autoradiography intracellular transport kinetics 9 photometry 9Kinetic studies of the secretory pathway have been limited by the methods commonly used, subcellular fractionation and electron microscopic autoradiography. These methods are well suited for identifying the organelles involved but are somewhat cumbersome for detailed kinetic studies. In the present paper, we describe widely applicable methods for quantitative light microscopic autoradiography. These methods have expedited the quantitative analysis of large numbers of autoradiograms, making possible detailed studies of secretory kinetics in the follicular epithelium of silkmoths. Studies using separation of cells from extracellular secreted products, plus preliminary electron microscope autoradiography, corroborated and extended the results of light microscope autoradiography. The major steps involved in secretion were identified and the duration of each step was evaluated. The follicular epithelium is well suited for studying secretory kinetics. It forms a monolayer around each oocyte and during the terminal pe-J. CELL BIOLOGY 9 The Rockefeller University Press -0021-9525/78/0701-013151.00 131

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Section: Quantitative Autoradiographymentioning

confidence: 99%

Secretory kinetics in the follicular cells of silkmoths during eggshell formation.

Blau¹,

Kafatos²

1978

The Journal of Cell Biology

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“…As demoastrated in earlier studies, the problems of track counting in SSNTD may be handled appropriately by electronic image analyzers Il, 4, 5]. Microphotome~ry appears to be a promising alternative and, in the past, proved successful for the quantitative determination of grair~ densities in film autoradiographs [2,3].The detector samples were prepared by coating microscope slides with a solution of cellulose nitrate (type 7E H P 3000, Dynamit Nobel, Germany), letting them dry overnight at room temperature ar~d for 3 days at 70 ° C. The detectors were exposed in contact with either planar standard c¢-sources [239Pu) electrodeposited on stainless steel planchets or with 20 ~zm thick femur sections of rats injected with 241Am. To vary the distance between source und detector, 1V[akrofol foils (Bayer, Germany) were placed between the detector and the s~andard ~-source.…”

mentioning

confidence: 97%

Microphotometry as a tool for automatic scanning of solid state nuclear track detectors

Kienzler

Polig

1975

Radiat Environ Biophys

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By use of a scanning microphotometer the light transmission with both dark field and bright field illumination was determined as a function of the alpha-track density on cellulose nitrate detectors. The so obtained calibration curves allow a rapit measurement of the number of tracks per field and, thus, of the dose rate. To illustrate the application of the method, a scan of a detector exposed on a bone section of a rat injected with 241Am is presented.

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Quantitative Autoradiography at the Cellular Level

Neuhoff

1973

Molecular Biology Biochemistry and Biophysics

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Auflichtphotometrie von Mikroautoradiogrammen für quantitative Einbaustudien an Einzelzellen

Cited by 14 publications

References 9 publications

Secretory kinetics in the follicular cells of silkmoths during eggshell formation.

Secretory kinetics in the follicular cells of silkmoths during eggshell formation.

Microphotometry as a tool for automatic scanning of solid state nuclear track detectors

Quantitative Autoradiography at the Cellular Level

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