Serial Sections for Electron Microscopy

Gay, Helen; Anderson, Thomas F.

doi:10.1126/science.120.3130.1071

Cited by 64 publications

(22 citation statements)

References 3 publications

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“…Because of the thin substrate, the process of making serial sections can be painstaking and is subject to tissue loss that poses a serious challenge for very large volume reconstructions (Gay and Anderson, 1954; Harris et al, 2006). With the introduction of high-performance field emission scanning electron microscopy (SEM) (Joy, 1991; Bogner et al, 2007), high quality images can be acquired from the surface of ultrathin sections thereby removing the need to mount sections on an electron-transmissive substrate.…”

Section: Introductionmentioning

confidence: 99%

Imaging ATUM ultrathin section libraries with WaferMapper: a multi-scale approach to EM reconstruction of neural circuits

Hayworth¹,

Morgan²,

Schalek³

et al. 2014

Front. Neural Circuits

252

198

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The automated tape-collecting ultramicrotome (ATUM) makes it possible to collect large numbers of ultrathin sections quickly—the equivalent of a petabyte of high resolution images each day. However, even high throughput image acquisition strategies generate images far more slowly (at present ~1 terabyte per day). We therefore developed WaferMapper, a software package that takes a multi-resolution approach to mapping and imaging select regions within a library of ultrathin sections. This automated method selects and directs imaging of corresponding regions within each section of an ultrathin section library (UTSL) that may contain many thousands of sections. Using WaferMapper, it is possible to map thousands of tissue sections at low resolution and target multiple points of interest for high resolution imaging based on anatomical landmarks. The program can also be used to expand previously imaged regions, acquire data under different imaging conditions, or re-image after additional tissue treatments.

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Section: Introductionmentioning

confidence: 99%

Imaging ATUM ultrathin section libraries with WaferMapper: a multi-scale approach to EM reconstruction of neural circuits

Hayworth¹,

Morgan²,

Schalek³

et al. 2014

Front. Neural Circuits

252

198

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“…Early electron microscopists quickly realized the value of obtaining serial sections to gather three-dimensional (3D) information 1 . The application of serial section technique to the study of 3D microcircuitry of the nervous system began more than 30 years ago 2 .…”

Section: Introductionmentioning

confidence: 99%

High-contrast en bloc staining of neuronal tissue for field emission scanning electron microscopy

et al. 2012

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Conventional heavy metal post staining methods on thin sections lend contrast but often cause contamination. To avoid this problem, we tested several en bloc staining techniques to contrast tissue in serial sections mounted on solid substrates for examination by Field Emission Scanning Electron Microscope (FESEM). Because FESEM section imaging requires that specimens have higher contrast and greater electrical conductivity than transmission electron microscope (TEM) samples, our technique utilizes osmium impregnation (OTO) to make the samples conductive while heavily staining membranes for segmentation studies. Combining this step with other classic heavy metal en bloc stains including uranyl acetate, lead aspartate, copper sulfate and lead citrate produced clean, highly contrasted TEM and SEM samples of insect, fish, and mammalian nervous system. This protocol takes 7–15 days to prepare resin embedded tissue, cut sections and produce serial section images.

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“…Blocks of embedded cells were prepared for serial sectioning as described by Fahrenbach (1984). Sections 70-80 nm thick were transferred to carbon-coated, slotted grids (Ted Pella, Redding, CA), as described by Gay and Anderson (1954). These preparations are not suitable for immunolabeling.…”

Section: Preparation and Purification Of Primary Antibodiesmentioning

confidence: 99%

Characterization of the Saccharomyces Golgi complex through the cell cycle by immunoelectron microscopy.

Preuss

Mulholland

Franzusoff

et al. 1992

MBoC

263

221

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The membrane compartments responsible for Golgi functions in wild-type Saccharomyces cerevisiae were identified and characterized by immunoelectron microscopy. Using improved fixation methods, Golgi compartments were identified by labeling with antibodies specific for alpha 1-6 mannose linkages, the Sec7 protein, or the Ypt1 protein. The compartments labeled by each of these antibodies appear as disk-like structures that are apparently surrounded by small vesicles. Yeast Golgi typically are seen as single, isolated cisternae, generally not arranged into parallel stacks. The location of the Golgi structures was monitored by immunoelectron microscopy through the yeast cell cycle. Several Golgi compartments, apparently randomly distributed, were always observed in mother cells. During the initiation of new daughter cells, additional Golgi structures cluster just below the site of bud emergence. These Golgi enter daughter cells at an early stage, raising the possibility that much of the bud's growth might be due to secretory vesicles formed as well as consumed entirely within the daughter. During cytokinesis, the Golgi compartments are concentrated near the site of cell wall synthesis. Clustering of Golgi both at the site of bud formation and at the cell septum suggests that these organelles might be directed toward sites of rapid cell surface growth.

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Serial Sections for Electron Microscopy

Cited by 64 publications

References 3 publications

Imaging ATUM ultrathin section libraries with WaferMapper: a multi-scale approach to EM reconstruction of neural circuits

Imaging ATUM ultrathin section libraries with WaferMapper: a multi-scale approach to EM reconstruction of neural circuits

High-contrast en bloc staining of neuronal tissue for field emission scanning electron microscopy

Characterization of the Saccharomyces Golgi complex through the cell cycle by immunoelectron microscopy.

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