CIL:12936, Rattus rattus, glandular epithelial cell, milk secreting cell, mammary alveolar cell. In Cell Image Library ...

Tissue was processed for immunoelectron microscopy using a modified Tokuyasu method. Briefly, minced tissue was fixed in 4% paraformaldehyde containing 5% sucrose and 100 mM HEPES and infiltrated with PBS containing 2.1 M sucrose over 10 h, with repeated solution changes. Fixed tissue was transferre...

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Bibliographic Details
Main Authors: Ladinsky, Mark, Howell, Kathryn E.
Format: Dataset
Language:unknown
Published: UC San Diego Library Digital Collections 2021
Subjects:
Online Access:https://dx.doi.org/10.6075/j09w0dnp
https://library.ucsd.edu/dc/object/bb6799050m
Description
Summary:Tissue was processed for immunoelectron microscopy using a modified Tokuyasu method. Briefly, minced tissue was fixed in 4% paraformaldehyde containing 5% sucrose and 100 mM HEPES and infiltrated with PBS containing 2.1 M sucrose over 10 h, with repeated solution changes. Fixed tissue was transferred to an aluminum cryosectioning stub (Ted Pella, Inc., Redding, CA) and immediately frozen in liquid nitrogen. Semithin (90 nm) cryosections were cut at -110C with an UltraCut UCT/FCS cryomicrotome (Leica), using a diamond knife (Diatome) and transferred to a Formvar-coated, carbon-coated, glow-discharged 100-mesh copper-rhodium electron microscopy grid. Following blocking of nonspecific antibody binding sites with 10% calf serum in PBS, the sections were labeled by sequential incubation with antibodies to adipophilin (guinea pig anti-adipophilin) and TGN38 (mouse monoclonal 2F7) and colloidal gold-conjugated secondary antibodies (15 nm anti-mouse, 10 nm anti-guinea pig; Ted Pella Inc., Redding, CA) and then ... : Preparation: cryostat-sectioned tissue; formaldehyde fixed tissue; embedded tissue; methylcellulose embedded Relation to intact cell: sectioned tissue Item type: recorded image Imaging mode: transmission electron microscopy (TEM); illumination by electrons; detection of electrons Parameter imaged: electron density Source of contrast: differences in adsorption or binding of stain; distribution of a specific protein Visualization methods: gold; uranyl salt; primary antibody plus labeled secondary antibody Processing history: unprocessed raw data Data qualification: Raw;spatialmeasurements;intensitiesquantitation ...