Scanning microscopy. Supplement最新文献_第3页

Aspects of cryofixation and cryosectioning for the observation of bulk biological samples in the hydrated state by cryoelectron microscopy. 低温电子显微镜观察水合状态下大块生物样品的冷冻固定和冷冻切片。

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

K Richter

引用次数: 0

Electro-optical imaging of F-actin and endoplasmic reticulum in living and fixed plant cells. 活的和固定的植物细胞中f -肌动蛋白和内质网的光电成像。

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

N S Allen, M N Bennett

{"title":"Electro-optical imaging of F-actin and endoplasmic reticulum in living and fixed plant cells.","authors":"N S Allen, M N Bennett","doi":"","DOIUrl":"","url":null,"abstract":"Confocal and video micrographs of living and fixed alfalfa roots, onion epithelial and pear pollen cells illustrate the architecture of the cytoskeleton and endoplasmic reticulum in plant cells. Fixation of plant tissues to preserve cytoplasmic structure poses special problems. When possible, emphasis should be placed on the imaging of structures in stained living cells over time. The early events that occur when Nod factors or bacteria elicit nodule formation in alfalfa roots will illustrate several approaches to plant cell fixation, staining and imaging. The first observable events after Nod factor stimulation occur in root hairs and are changes in rates of cytoplasmic streaming, nuclear movements, and changes in the shape of the vacuole. Within ten minutes, the endoplasmic reticulum shifts position towards the tip of the root hair. For comparison, the endoplasmic reticulum localization in pollen tubes and onion epithelial cells will be illustrated. The actin cytoskeleton undergoes a series of changes over a twelve hour period. These changes in the cytoskeleton are spatially and temporally correlated with the observed growth changes of the root hairs. This dynamic change of the actin filament and endoplasmic reticulum and associated secretory vesicles in these root hairs suggests a mechanism for the observed root hair growth changes.","PeriodicalId":77379,"journal":{"name":"Scanning microscopy. Supplement","volume":"10 ","pages":"177-86; discussion 186-7"},"PeriodicalIF":0.0,"publicationDate":"1996-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20522142","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

Atomic force microscopy of DNA, nucleoproteins and cellular complexes: the use of functionalized substrates. DNA、核蛋白和细胞复合体的原子力显微镜:功能化底物的使用。

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

Y L Lyubchenko, R E Blankenship, A A Gall, S M Lindsay, O Thiemann, L Simpson, L S Shlyakhtenko

{"title":"Atomic force microscopy of DNA, nucleoproteins and cellular complexes: the use of functionalized substrates.","authors":"Y L Lyubchenko, R E Blankenship, A A Gall, S M Lindsay, O Thiemann, L Simpson, L S Shlyakhtenko","doi":"","DOIUrl":"","url":null,"abstract":"Progress towards rapid and simple characterization of biomolecular samples by scanning probe microscopy is impeded mainly by limitations of the current approach to sample preparation. We are working on approaches based on chemical functionalization of mica. Treatment of mica with aminopropyltriethoxy silane (APTES) makes the surface positively charged (AP-mica) and able to hold DNA in place for imaging, even in water. We have shown that AP-mica is an appropriate substrate for numerous nucleoprotein complexes as well. The AFM images of the complex of DNA with RecA protein are stable and indicate a structural periodicity for this filament. AP-mica holds strongly such large DNA complexes as kinetoplast DNA (kDNA) and is an appropriate substrate for their imaging with AFM. We have further develop this approach for making hydrophobic substrates. Silylation of mica surface with hexamethyldisilazane (Me-mica) allowed us to get AFM images of chlorosomes, an antenna complex isolated from green photosynthetic bacteria. Me-mica may be converted into a positively charged substrate after treatment with water solutions of tetraethylammonium bromide or cetyltrimethylammonium bromide. These activated surfaces show high activity towards binding the DNA molecules.","PeriodicalId":77379,"journal":{"name":"Scanning microscopy. Supplement","volume":"10 ","pages":"97-107; discussion 107-9"},"PeriodicalIF":0.0,"publicationDate":"1996-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20521594","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

Covalent labeling of proteins with fluorescent compounds for imaging applications. 荧光化合物在蛋白质成像应用中的共价标记。

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

D R Swartz

引用次数: 0

Specimen preparation of the human cerebellar cortex for scanning electron microscopy using a t-butyl alcohol freeze-drying device. 用丁醇冷冻干燥装置制备用于扫描电子显微镜的人小脑皮层标本。

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

T Hojo

引用次数: 0

In vitro systems and cultured cells as specimens for X-ray microanalysis. 体外系统和培养细胞作为x射线微量分析的标本。

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

G M Roomans, J Hongpaisan, Z Jin, A C Mörk, A Zhang

{"title":"In vitro systems and cultured cells as specimens for X-ray microanalysis.","authors":"G M Roomans, J Hongpaisan, Z Jin, A C Mörk, A Zhang","doi":"","DOIUrl":"","url":null,"abstract":"In vitro systems and cultured cells are recognized as useful systems in many areas of biomedical research, including X-ray microanalysis. To be reliable, in an vitro system should have an elemental composition close to that of the tissue in situ, react in the same way to stimuli, and retain the in situ regulation of ion transport. In the present paper, four of the most commonly used in vitro systems will be reviewed: incubated tissue slices (liver and pancreas), isolated glands (submandibular gland acini, sweat glands), primary cell cultures (sweat glands, endometrium), and cell lines (the colon cancer cell line T84, immortalized sweat gland cells). Incubation of tissue slices of liver in Krebs-Ringers buffer caused a significant increase in Na and Cl and a decrease in K. Initially, these changes were also observed in the pancreas, but here the values gradually returned to normal. Isolated submandibular gland acini, and isolated sweat gland ducts and coils react in a similar way to stimulation as their in situ counterparts. In primary cultures of coil cells, however, part of the cell population acquires different ion transport characteristics. Technically simplest is the use of cell lines originating from cancer cells (e.g., the T84 cell line) and immortalized cell lines. X-ray microanalysis not only confirms data on ion transport obtained with other techniques, but adds the possibility to investigate the presence of subpopulations within a culture.","PeriodicalId":77379,"journal":{"name":"Scanning microscopy. Supplement","volume":"10 ","pages":"359-70; discussion 370-3"},"PeriodicalIF":0.0,"publicationDate":"1996-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20522054","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

Nucleic acid detection by in situ molecular immunogold labeling procedures. 核酸检测采用原位分子免疫金标程序。

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

M Thiry

引用次数: 0

Preparation of plasmid DNA in transfection complexes for fluorescence and electron spectroscopic imaging. 质粒DNA转染复合物的制备及其荧光和电子光谱成像。

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

M Malecki

{"title":"Preparation of plasmid DNA in transfection complexes for fluorescence and electron spectroscopic imaging.","authors":"M Malecki","doi":"","DOIUrl":"","url":null,"abstract":"The aim of this project was to develop procedures necessary to study mechanisms of receptor mediated gene transfer by means of integrated microscopy. Plasmid DNA was incorporated into a transfection complex consisting of poly(L)lysine and transferrin to which the nuclear localization signal was conjugated. This complex was presented to cultured glioma cells. Preparation of the transfected DNA for imaging was pursued by two methods. In the first method tetramethylrhodamine, nanogold, and ferritin were linked through streptavidin to the biotinylated plasmid DNA. Trafficking of the fluorescent derivatives was studied in living cells with fluorescence microscopy. Then, selected cells were rapidly cryo-immobilized. Ultra-structural distribution of the transfected DNA was imaged with energy filtering transmission electron microscopy. In the second method, the unmodified transfected DNA was detected in cryo-immobilized cells by in situ polymerase chain reaction and in situ hybridization. For laser scanning fluorescence microscopy probes were labeled with tetramethylrhodamine. For ultrastructural analysis by electron spectroscopic imaging, probes containing incorporated digoxigenin were labeled with anti-digoxigenin boronated antibodies. Based upon the developed procedures, it has been demonstrated that the presence of the nuclear localization signal in the transfection complex resulted in rapid nuclear import of the transfected DNA.","PeriodicalId":77379,"journal":{"name":"Scanning microscopy. Supplement","volume":"10 ","pages":"1-16"},"PeriodicalIF":0.0,"publicationDate":"1996-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20521686","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

Cytoskeleton architecture of C6 rat glioma cell subclones whole mount electron microscopy and immunogold labeling. C6大鼠胶质瘤细胞亚克隆的细胞骨架结构及免疫金标记。

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

W Bohn, D Etzrodt, R Foisner, G Wiche, P Traub

引用次数: 0

Comparative scanning, transmission and atomic force microscopy of the microtubular cytoskeleton in fenestrated liver endothelial cells. 开窗肝内皮细胞微管细胞骨架的扫描、透射和原子力显微镜对比研究。

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

F Braet, R De Zanger, W Kalle, A Raap, H Tanke, E Wisse

{"title":"Comparative scanning, transmission and atomic force microscopy of the microtubular cytoskeleton in fenestrated liver endothelial cells.","authors":"F Braet, R De Zanger, W Kalle, A Raap, H Tanke, E Wisse","doi":"","DOIUrl":"","url":null,"abstract":"Endothelial fenestrae control the exchange of fluids, solutes and particles between the sinusoidal lumen and the microvillous surface of the parenchymal cells. Fenestrae have a critical dimension in the order of 150-200 nm, making it necessary to use microscopes with a resolution better than the light microscope. Comparative whole-mount preparations of isolated, purified and cultured rat liver sinusoidal endothelial cells (LEC) were studied by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and atomic force microscopy (AFM). Examination of detergent-extracted LEC by SEM and TEM shows an integral cytoskeleton: sieve plates are delineated by a sieve plate-associated cytoskeleton ring and fenestrae by a fenestrae-associated cytoskeleton ring. By using microtubule altering agents we could demonstrate: (1) the architectural role of microtubules in arranging fenestrae, (2) the existence of a population of microtubules resistant against low temperature and colchicine, (3) the ability of LEC to shift the microtubule assembly-disassembly steady state under various conditions, (4) and the necessity of an intact microtubular cytoskeleton to support the increase in the number of fenestrae after cytochalasin B. Topographical examinations of AFM images revealed that sieve plates are delineated by elevated borders, probably projections of the underlying tubular cytoskeleton.","PeriodicalId":77379,"journal":{"name":"Scanning microscopy. Supplement","volume":"10 ","pages":"225-35; discussion 235-6"},"PeriodicalIF":0.0,"publicationDate":"1996-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20522146","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