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*CIL – Cell Image Library accession number. Please use this to reference an image.

CIL:50900*  Cite 

Wild type Saccharomyces cerevisiae cells were high pressure frozen and embedded in HM20 resin. Samples were then stained for Nuclear Pore Complex (NPC) assosiated proteins with the monoclonal antibody ab24609 (Abcam) and visualized through transmission electron microscopy (TEM). The images show the nucleus of the cells and the black arrow indicates nuclear envelope budding events. The black gold particles represent the presence of NPC proteins.

Technical Details

Wild type cells were cultured in YPD medium and high pressure frozen in a Wohlwend Compact 3 machine, followed by a short freeze substitution protocol in 2% Uranyl acetate and embedded in HM20 resin. For the immuno-EM, samples were blocked for 2 hours in 1% BSA with 0.1% fish skin gelatine. A 1:50 dilution of the primary antibody ab24609 was used and a 1:150 dilution of the secondary antibody rabbit anti-mouse immunoglobulins (E0433, Agilend/Dako). Another incubation with a 1:70 diluted 10 nm gold-conjugated protein A (E1808, CMC UMC Utrecht) was followed. Finally, samples were post fixed in 2.5% glutaraldehyde for 20 minutes. Sections of 70 nm thickness were then contrast stained with 2% Uranyl acetate and Reynold's lead citrate. Pictures were aquired in a Tecnai T12 TEM with a Ceta CMOS 16M camera.

Biological Sources
NCBI Organism Classification
Saccharomyces cerevisiae (baker's yeast, budding yeast)" Caenorhabditis elegans (roundworm, nematode) Human mast cell line (HMC-1)
Cell Type
Mixed population of S. cerevisiae cells
Cellular Component
Biological Context
Biological Process
Nuclear envelope budding and Nuclear Pore Complex assosiated proteins localization
Johanna Höög
Dimitra Panagaki
Jacob Croft
Digital Object Identifier (DOI)
Archival Resource Key (ARK)
Grouping This image is part of a group.
Image Mode
transmission electron microscopy (TEM)
Visualization Methods
Stain with Uranyl acetate and Lead citrate