Quantitative evaluation of autoradiographs by X‐ray spectroscopy

Hodges, Gisèle M.; Muir, M. D.

doi:10.1111/j.1365-2818.1975.tb04014.x

Cited by 13 publications

(6 citation statements)

References 8 publications

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“…Thus, it was demonstrated that X-ray microanalysis by TEM mode was useful in quantifying silver grains in EMRAG instead of visual grain counting. In the literature, Hodges and Muir [10] were the first to quantify silver content on EMRAG obtained from cultured cells labeled with 3 H-thymidine by scanning electron microscopy (SEM) equipped with EDX, but they did so by observing the cell surface without confirming the nuclei and cell organelles surrounded with cytoplasm. Our reports [27,30,32] were the first to quantify the silver grains by TEM mode observing both the silver grains and the nuclei and cell organelles.…”

Section: Silver Grains In Radioautographs (Ag)mentioning

confidence: 99%

The Utility Value of High Voltage Electron Microscopy for X-Ray Microanalysis

Nagata

2003

Acta Histochem. Cytochem

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X-ray microanalysis is a useful technique to qualify and quantify basic elements in biological specimens. This article reviews the principles and techniques for applying intermediate high voltage electron microscopy to Xray microanalysis of various elements in biological specimens developed in our laboratory since the late 20th century. We first quantified the endproducts of histochemical reactions such as Ag in radioautographs, Ce in acid phosphatase reaction and Au in colloidal gold immunostaining using semi-thin sections by high voltage electron microscopy at 300-400 kV. We then analyzed various trace elements such as Zn, Ca, and S, which originally existed in the cytoplasmic matrix or cell organelles of various cells in different organs, and some absorbed elements introduced by experimental administration into cells and tissues such as Al, using both conventional chemical fixation and cryofixation followed by cryo-sectioning, freeze-drying, or freeze-substitution. As a result, we showed that the peak to background (P/B) ratios of all the elements analyzed had high P/B ratios at 300-400 kV. It was concluded that X-ray microanalysis using semi-thin sections by high voltage electron microscopy is of great utility for quantifying trace elements in biological specimens.

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Section: Silver Grains In Radioautographs (Ag)mentioning

confidence: 99%

The Utility Value of High Voltage Electron Microscopy for X-Ray Microanalysis

Nagata

2003

Acta Histochem. Cytochem

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“…10a and lob) as a consequence of the.high atomic number contrast of the heavy metal against the lower contrast of the biological material (Hodges & Muir, 1976;Bowen et al, 1983). Where necessary, further identification or quantification of silver grains should be possible by X-ray analysis as previously discussed (Hodges & Muir, 1975;Junger & Bachmann, 1980). By immunofluorescence, the He117 fibroblasts displayed the characteristic fibrillar network of extracellular fibronectin using either monospecific or monoclonal antifibronectin antibodies (Fig.…”

Section: Observations Of Emulsion-coated [3h]mentioning

confidence: 97%

“…9a, b and 10a, b). Where necessary, further identification or quantification of silver grains should be possible by X-ray analysis as previously discussed (Hodges & Muir, 1975;Junger & Bachmann, 1980). The appearance of the silver grains was not always easy to recognize when samples were examined in the secondary electron imaging (SEI) mode and could be confused with the microvillous configurations of the RT4 cell surface.…”

Section: Sem Analysis Of Autoradiogramsmentioning

confidence: 99%

A methodological basis for SEM autoradiography: biosynthesis and radioligand binding

Cheng

Hodges

Trejdosiewicz

1985

Journal of Microscopy

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SUMMARY A method is described for scanning electron microscope (SEM) autoradiography whereby preservation of high resolution cell surface details is retained together with degelatination of the emulsion without gross loss or redistribution of silver grains. This method should provide a convenient medium‐sized marker for SEM (using secondary, backscattered electron and X‐ray imaging) topographic studies of biosynthesized molecules, and of cell surface receptors and antigens, using indirect or direct labelling procedures with radio‐labelled ligands.

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“…A related field, the exploration of which is only beginning, is the use of cell surface markers incorporating distinctive elements (Baccetti & Burrini, 1977;Hoyer et al, 1979) which can be detected by X-ray microanalysis, rather than the more usual fluorescent or enzyme markers. X-ray microanalysis has also been proposed for the quantification of silver grains on autoradiographs (Hodges & Muir, 1975) but, again, this application has not yet been much employed.…”

Section: Diffusible Ionsmentioning

confidence: 99%

X-ray microanalysis: a histochemical tool for elemental analysis

1983

Histochem J

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The interactions of electrons with specimens Types of interactionsWhen electrons strike an object, as in an electron microscope, a number of interactions occur that can be, and are, used to give information about the specimen (Table 1). The various types of interactions of electrons that are used to produce images in scanning or transmission electron microscopes are familiar to most biologists, in the form of electron micrographs. The density of a transmission electron micrograph is dependent on the mass of the specimen at that point; because of this, staining and histochemical methods have been developed for electron microscopy using heavy metals. In scanning electron microscopy, the generation of back-scattered electrons is also dependent on atomic number. The information that can be obtained about the composition of the specimen in these ways is, however, extremely crude compared with that obtainable by some other methods. Among these other methods is X-ray microanalysis (also known as electron probe microanalysis) which is foremost among such techniques at present for ease of application and interpretation, and availability of commercial equipment. In X-ray microanalysis, the electrons striking the specimen excite X-rays characteristic of the elements in the specimen. X-ray microanalysis is thus a form of elemental analysis. It has been applied to a wide variety of biological problems, and these will be considered later in the section entitled Some biological applications of X-ray microanalysis. The production of X-rays by electronsWhen an electron of sufficient energy passes through the cloud of electrons surrounding an atomic nucleus, it may cause the displacement of an inner orbital electron to an orbit of higher energy. In the process the primary electron is inelastically scattered; that is, it

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Quantitative evaluation of autoradiographs by X‐ray spectroscopy

Cited by 13 publications

References 8 publications

The Utility Value of High Voltage Electron Microscopy for X-Ray Microanalysis

The Utility Value of High Voltage Electron Microscopy for X-Ray Microanalysis

A methodological basis for SEM autoradiography: biosynthesis and radioligand binding

X-ray microanalysis: a histochemical tool for elemental analysis

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