Cathodoluminescence applied to immunofluorescence: Present state and improved technical prospects by prism spectrometer light selection

Cavellier, J. F.; Berry, J. P.; Lagrue, G

doi:10.1007/bf00492667

Cited by 10 publications

(5 citation statements)

References 14 publications

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“…Therefore, a new system which can effectively collect CL signal and minimize the beam effect has been required. Initially, the CL collection system consisted of a light pipe or lens and PMT (Cavellier et al, 1978;Schmidt et al, 1975); thereafter various concave mirrors, such as parabolic (Bond et al, 1974;Muir et al, 1971) and hemispheric mirrors (Jugde et al, 1974), were introduced in order to achieve a large solid angle of CL emission. Hörl (1972) adopted an ellipsoidal mirror, which we also introduced to maintain a large solid angle and minimize transmission losses of CL.…”

Section: Discussionmentioning

confidence: 99%

“…Although CL is essentially a similar phenomenon to conventional fluorescence elicited under ultraviolet light, it has some intrinsic advantages over the latter (Bröcker and Pfefferkorn, 1979;Pfefferkorn et al, 1980): (1) CL can be amplified electronically and measured quantitatively; (2) the depth of focus and the resolution in the CL mode is much higher than in the conventional fluorescence microscope; and (3) some substances exhibit luminescence only under electron irradiation. Although many attempts (Basu, 1983;Bond et al, 1974;Boyde and Reid, 1983;Bröcker and Pfefferkorn, 1979;Cavellier and Berry, 1985;Cavellier et al, 1978;De Mets, 1974;De Mets and Lagasse, 1971;Herbst and Hoder, 1978;Hörl, 1972Hörl, , 1978Hurter et al, 1981;Pearce and Hays, 1966;Schmidt et al, 1975;Soni et al, 1975) were made to utilize CL in biology, its applications have been limited because CL from organic chemicals is a very weak signal and easily fades under high beam current (De Mets and Lagasse, 1971).…”

Section: Introductionmentioning

confidence: 97%

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Analytical color fluorescence electron microscopy of adrenal cortex

Nakano

Koike

Ogawa

1997

Microsc. Res. Tech.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 97%

Analytical color fluorescence electron microscopy of adrenal cortex

Nakano

Koike

Ogawa

1997

Microsc. Res. Tech.

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“…In the early days of CL research, the CL collection system consisted of a light pipe or lens and PMT (8,32), and so did our system in its early development. In order to achieve a large solid angle of CL emission, various-shaped concave mirrors such as a parabolic (3,24) and a hemispheric mirror (20) were introduced.…”

Section: Postnatal Developmentmentioning

confidence: 99%

Application of analytical color fluorescence electron microscopy to biomedical field: I. Vitamin A ester in rat retina.

Nakano

Fujimoto

Koike

et al. 1990

Acta Histochem. Cytochem

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An analytical color fluorescence electron microscope (ACFEM) based on a high-resolution scanning electron microscope has been developed. The ACFEM enables us not only to detect cathodoluminescence (CL), which is a weak luminescence signal under electron beam bombardments, as color images, but also to analyze CL spectra. A cryo-SEM method was introduced to prevent beam effect on biological specimens. In experiments 1 and 2, we observed adult rat retinas under different conditions: hyper-and hypovitaminosis A and light and dark adaptation, which revealed that the distribution of vitamin A ester and its change under these conditions could be detected in situ by the ACFEM. In experiment 3, postnatal development of rat retina was observed under the ACFEM up to 3 weeks after birth. The retinal pigment epithelial cells of new born rats were already functioning as vitamin A storing cells. On the other hand, vitamin A ester in the developing outer segment first appeared on the 13th postnatal day, which suggests a correlation to the development of visual function. These results show that CL analysis by the ACFEM is a simple and effective new method in the field of histo-and cytochemistry.In scanning electron microscopy, several signals can be utilized such as secondary electrons, backscattered electrons, transmitted electrons, Auger's electrons and Xrays. At the same time, cathodoluminescence (CL) is emitted as a light signal in the visible, ultraviolet and infrared regions when substances are bombarded with an electron beam (30). Although the CL phenomenon is essentially similar to fluorescence and phosphorescence elicited by ultraviolet light, it has some intrinsic advantages over them (6, 30): 1) CL can be amplified electronically and measured quantitatively, 2) the depth of focus and the resolution in the CL mode is higher than in the conventional fluorescence microscope, 3) some substances exhibit luminescence only under electron irradiation. In the industrial field, the CL technique has been introduced as a practically valuable tool for analyzing wide band-gap material such as diamonds (26). However, in the biomedical field, although many attempts have been made to Correspondence to: Tohru Nakano,

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“…In the early days, the CL-collecting system consisted of a light pipe or lens and PMT (10,33). We also used such a detector system and investigated various sorts of CL-emitting substances in the previous paper (14).…”

Section: Detector Systemmentioning

confidence: 99%

Color cathodoluminescence images and cathodoluminescence spectra analysis of biological materials.

Itoh

Fujimoto

Koike

et al. 1986

Acta Histochem. Cytochem

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In an attempt to detect cathodoluminescence (CL), an Akashi scanning electron microscope (SEM) ISI-DS130 was remodeled into a cathodoluminescence electron microscope (CEM or fluorescence electron microscope FEM). A newly shaped mirror, combining three half ellipsoidal mirrors, was located over the specimen holder of the top stage for color CL use. The bottom stage was modified with a half ellipsoidal mirror, photomultiplier tube (PMT) and monochrometer, so that not only CL images, but also CL spectra could be obtained.An effective specimen-preparation method, known as the simple cryo-SEM method (24), was devised for CL observation.With these instruments and methods, color CL images and CL spectra of the biological substances were analyzed.Ito cells of vitamin A (vit A)-loaded rat liver and zona fasciculata cells of rat adrenal cortex emitted intense CL from their lipid droplets (LD). In order to identify the origin of this CL emission, analysis of the color CL images and the CL spectra were available.Cathodoluminescence (CL) is a light emission, in the visible, near ultraviolet and infrared regions under the bombardment of an electron beam (31). CL phenomenon is similar to fluorescence or phosphorescence excited by ultraviolet light, but it has some intrinsic advantages over the fluorescence as described by Brocker et al. (6,31) : 1) CL can be amplified and measured quantitatively, 2) The depth of focus and the resolution in the CL mode are higher than in the fluorescence microscope. 3) Some substances exhibit luminescence only under electron irradiation. Furthermore, CL of organic chemicals stems from loosely bound it electrons; it shows the information of the chemical bond. Just as secondary electrons (SE), backscattered electrons, Auger electrons and X-rays are often used for observing or analysing substances under scanning electron microscope (SEM), so CL should become an effective analytical tool.Although CL studies as an electron probe can be traced to 1952, the CL applications to biological and medical fields have not made remarkable advances.

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Cathodoluminescence applied to immunofluorescence: Present state and improved technical prospects by prism spectrometer light selection

Cited by 10 publications

References 14 publications

Analytical color fluorescence electron microscopy of adrenal cortex

Analytical color fluorescence electron microscopy of adrenal cortex

Application of analytical color fluorescence electron microscopy to biomedical field: I. Vitamin A ester in rat retina.

Color cathodoluminescence images and cathodoluminescence spectra analysis of biological materials.

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