Scanning microscopy. Supplement最新文献_第4页

Pre-embedding staining of single muscle fibers for light and electron microscopy studies of subcellular organization. 单个肌肉纤维的预包埋染色用于亚细胞组织的光学和电子显微镜研究。

Scanning microscopy. Supplement Pub Date : 1996-01-01

E Ralston, T Ploug

引用次数: 0

Immunocytochemistry by electron spectroscopic imaging using well defined boronated monovalent antibody fragments. 免疫细胞化学通过电子光谱成像使用明确的硼化单价抗体片段。

Scanning microscopy. Supplement Pub Date : 1996-01-01

M M Kessels, B Qualmann, W D Sierralta

{"title":"Immunocytochemistry by electron spectroscopic imaging using well defined boronated monovalent antibody fragments.","authors":"M M Kessels, B Qualmann, W D Sierralta","doi":"","DOIUrl":"","url":null,"abstract":"Contributing to the rapidly developing field of immunoelectron microscopy a new kind of markers has been created. The element boron, incorporated as very stable carborane clusters into different kinds of peptides, served as a marker detectable by electron spectroscopic imaging (ESI)--an electron microscopic technique with high-resolution potential. Covalently linked immunoreagents conspicuous by the small size of both antigen recognizing part and marker moiety are accessible by using peptide concepts for label construction and their conjugation with Fab' fragments. Due to a specific labeling of the free thiol groups of the Fab' fragments, the antigen binding capacity was not affected by the attachment of the markers and the resulting immunoprobes exhibited an elongated shape with the antigen combining site and the label located at opposite ends. The labeling densities observed with these reagents were found to be significantly higher than those obtained by using conventional colloidal gold methods. Combined with digital image processing and analysis systems, boron-based ESI proved to be a powerful approach in ultrastructural immunocytochemistry employing pre- and post-embedding methods.","PeriodicalId":77379,"journal":{"name":"Scanning microscopy. Supplement","volume":"10 ","pages":"327-44"},"PeriodicalIF":0.0,"publicationDate":"1996-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20522051","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Optical methods for imaging ionic activities. 离子活性成像的光学方法。

Scanning microscopy. Supplement Pub Date : 1994-01-01

R B Moreton

{"title":"Optical methods for imaging ionic activities.","authors":"R B Moreton","doi":"","DOIUrl":"","url":null,"abstract":"Optical fluorescence is characteristic of molecules and their environment, and dyes can be made whose fluorescence is altered by reversible binding to specific ions. By introducing these into the cytosol, fluorescence microscopy can be used to form dynamic images of ionic activities in living cells under experimental manipulation. Optical fluorescence spectra are broad-band, and if specific ion binding alters the wavelength of maximal excitation or emission, quantitative measurements can be made from the ratio of images taken at two different wavelengths, eliminating errors due to spatial variations in dye concentration and optical path-length. This method is analogous to continuum normalisation in X-ray microanalysis, and is implemented using a sensitive video camera and computer processing digitised images. Fluorescent indicators exist for calcium, magnesium, hydrogen, sodium, zinc and chloride ions. Most imaging work has been on calcium, which is important in many cell signalling processes, and several calcium indicators are available with different spectral properties. Spatial resolution is limited to a few micron by out-of-focus blur, but repeated images can be captured with a time resolution as low as 200 msec, and by using dyes with high binding affinity, detection limits can be lower than by X-ray methods. There is a large and fast-growing literature of applications to many plant and animals cell-types.","PeriodicalId":77379,"journal":{"name":"Scanning microscopy. Supplement","volume":"8 ","pages":"371-90"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18643236","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Image-EELS: a synthesis of energy-loss analysis and imaging. 图像- eels:能量损失分析和成像的综合。

Scanning microscopy. Supplement Pub Date : 1994-01-01

K H Körtje

{"title":"Image-EELS: a synthesis of energy-loss analysis and imaging.","authors":"K H Körtje","doi":"","DOIUrl":"","url":null,"abstract":"Two different modes of energy-filtering transmission electron microscopy (EFTEM) are often used for element microanalysis: electron energy-loss spectroscopy (EELS) and electron spectroscopic imaging (ESI). A new approach was developed which we call Image-EELS. This procedure was realized with the commercially available standard equipment of the energy-filtering transmission microscope CEM 902 (Zeiss, Germany). A series of energy-filtered images is recorded with ESI at many different energy losses. In a second step the intensity of selected objects is measured for each energy loss and plotted as a function of the energy loss, that means as an EELS spectrum. This method increases the sensitivity of EELS analysis, especially for very small and irregular objects, because the lateral resolution is enhanced and the noise is suppressed by the integration of many pixels belonging to one type of object. Many spectra can be calculated from one image series, enabling the comparison of spectra from different objects. Selected images from the series can be used for ESI elemental mapping, so that errors and limits in the different mapping procedures can be detected. Image-EELS is a synthesis of EELS and ESI and as such it constitutes a considerable progress for element microanalysis with EFTEM, not only for biological objects.","PeriodicalId":77379,"journal":{"name":"Scanning microscopy. Supplement","volume":"8 ","pages":"277-87;discussion 287-8"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18643290","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The principles of proton probe microanalysis in biology. 生物学中质子探针微量分析原理。

Scanning microscopy. Supplement Pub Date : 1994-01-01

G J Legge, M Cholewa

引用次数: 0

Measurement of subcellular Ca2+ redistribution in cardiac muscle in situ: time resolved rapid freezing and electron probe microanalysis. 心肌亚细胞Ca2+再分布原位测量:时间分辨快速冷冻和电子探针微量分析。

Scanning microscopy. Supplement Pub Date : 1994-01-01

M Bond, M D Schluchter, E Keller, C S Moravec

{"title":"Measurement of subcellular Ca2+ redistribution in cardiac muscle in situ: time resolved rapid freezing and electron probe microanalysis.","authors":"M Bond, M D Schluchter, E Keller, C S Moravec","doi":"","DOIUrl":"","url":null,"abstract":"To directly assess the physiological roles of sarcoplasmic reticulum (SR) and mitochondria (MT), we have utilized energy dispersive electron probe microanalysis (EPMA) on ultrathin freeze-dried cryosections from isolated papillary muscles, rapidly frozen at precise time points of the contractile cycle. Using this approach, we can detect redistribution of subcellular Ca2+ during the cardiac contractile cycle. Changes in Ca2+ of less than 1.0 mmol/kg dry wt can be detected. By determining the variability of the Ca2+ measurements in preliminary experiments, we have also demonstrated that it is possible to optimize experimental design, i.e., to predict the number of animals per treatment group and the number of X-ray spectra per animal that are required in order to detect a specified Ca2+ difference. Quantitative EPMA of rapidly frozen contracting papillary muscle has also allowed us to correlate the Ca2+ content of SR and MT with the contractile state of the muscle. Our results show a decrease of 40% in the amount of Ca2+ stored in the junctional SR during a cardiac muscle twitch, thus providing direct evidence for a role of the SR as a primary site of Ca2+ release. In addition, we have demonstrated dissociation between MT Ca2+ uptake and activation of regulatory enzymes, such as pyruvate dehydrogenase, indicating that MT Ca2+ uptake is not required for activation of MT metabolism.","PeriodicalId":77379,"journal":{"name":"Scanning microscopy. Supplement","volume":"8 ","pages":"1-11"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18644779","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Light element X-ray microanalysis in biology. 生物学中的轻元素x射线微量分析。

Scanning microscopy. Supplement Pub Date : 1994-01-01

A T Marshall

{"title":"Light element X-ray microanalysis in biology.","authors":"A T Marshall","doi":"","DOIUrl":"","url":null,"abstract":"It is shown that both qualitative and quantitative light element X-ray microanalysis of biological samples is feasible. These analyses were carried out using ultrathin window (UTW) detectors. Quantitative analysis yields a total element analysis with H estimated by difference or \"guesstimated\". Comparison with calculated concentrations, or concentrations obtained by chemical analysis, shows that X-ray microanalysis of sections, by the peak to continuum ratio model, give sufficiently accurate results for biological purposes. The measurement of O concentrations to yield water content is carried out using x-ray imaging techniques, so that the distribution of heavier elements can be spatially related to water and dry mass distribution. Similarly light element and heavy/light element ratios are readily visualised by X-ray imaging. These ratios can indicate the subcellular distribution of different molecular species e.g., nitrogenous compounds such as urates. It is possible to derive quantitative images of water distribution in both sections and bulk samples. Comparisons of the same sample type both as frozen sections and frozen bulk samples show that the water estimates obtained by the two different analytical methods are similar. Oxygen analysis of C films at different specimen temperatures unequivocally reveals the temperature at which ice deposition on the specimen commences. This establishes safe conditions for reducing mass loss in model samples and freeze-dried sections to minimal levels and for avoiding artefactual oxygen analyses of both frozen-hydrated and freeze-dried sections.","PeriodicalId":77379,"journal":{"name":"Scanning microscopy. Supplement","volume":"8 ","pages":"187-99; discussion 199-201"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18642703","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Optimal freeze-drying of cryosections and bulk specimens for X-ray microanalysis. 用于x射线显微分析的冷冻切片和散装标本的最佳冷冻干燥。

Scanning microscopy. Supplement Pub Date : 1994-01-01

L Edelmann

引用次数: 0

Energy-filtering transmission electron microscopy as a tool for structural and compositional analysis of isolated ferritin particles. 能量过滤透射电子显微镜作为分离铁蛋白颗粒结构和成分分析的工具。

Scanning microscopy. Supplement Pub Date : 1994-01-01

A L Beckers, W C de Bruijn, J F Jongkind, M I Cleton-Soeteman, R P Apkarian, E S Gelsema

{"title":"Energy-filtering transmission electron microscopy as a tool for structural and compositional analysis of isolated ferritin particles.","authors":"A L Beckers, W C de Bruijn, J F Jongkind, M I Cleton-Soeteman, R P Apkarian, E S Gelsema","doi":"","DOIUrl":"","url":null,"abstract":"Structural and compositional analysis of isolated horse-spleen ferritin particles was performed by energy filtering transmission electron microscopy (EFTEM). Ferritin particles were collected in ultrathin (2 nm thick) chromium films and analyzed without any additional stain by electron energy-loss spectroscopy (EELS) for iron and carbon and by electron-spectroscopic imaging (ESI) for carbon. The ultrastructure of the proteinaceous shell of the ferritin particle, as obtained by the carbon net-intensity electron spectroscopical and carbon concentration-distribution images, was qualitatively compared to the structure as acquired by a negative-staining procedure. Quantitative analysis of the number of carbon atoms in the ferritin-shell proteins was carried out through an ESI-acquisition protocol and processing procedure with calibrated attenuation filters in the optical path to the TV camera. This procedure included images acquired with calibrated attenuation filters for the compensation of shading and the non-linear performance of the TV camera used in the analytical part of the procedure. A new ¿ESI-Spectra¿ program is proposed that allows element-related spectra to be generated at any place and with any frame size in a contrast-sensitive or other type of image present on the computer monitor screen.","PeriodicalId":77379,"journal":{"name":"Scanning microscopy. Supplement","volume":"8 ","pages":"261-74;discussion 274-5"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18643289","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Quantitative biological electron probe microanalysis with a wavelength dispersive spectrometer. 波长色散光谱仪定量生物电子探针微量分析。

Scanning microscopy. Supplement Pub Date : 1994-01-01

A Pogorelov, V Pogorelova, N V Repin, I Mizin

引用次数: 0