Alternate header for print version

Attribution Non-Commercial Share Alike:This image is licensed under a Creative Commons Attribution, Non-Commercial Share Alike License. View License Deed | View Legal Code
*CIL – Cell Image Library accession number. Please use this to reference an image.

CIL:13864*  Cite 

Cdc15-3HA (red) does not load onto spindle pole bodies in wild-type cells in metaphase as determined by spindle morphology (tubulin, green) and nuclear morphology (DAPI, blue). Control image for CIL# 13862, 13863 in which constitutive targeting of Tem1 to spindle pole bodies leads to premature Cdc15 loading. Image is Fig 3C, top panels, in J Cell Biol. (2011) 192: 599-614. Images in Fig 3 include CIL# 13862, 13863, 13864, 13865, 13866, 13867.

Technical Details

Cells (MATa cdc15::CDC15-3HA), grown in rich media with glucose, were fixed for 15 min in 3.7% formaldehyde and 0.1 M potassium phosphate buffer, pH 6.4. Cells were then washed twice with 0.1 M potassium phosphate buffer, pH 6.4, and resuspended in 1.2 M sorbitol in 0.12 M K2HPO4/0.033 M citric acid, pH 5.9. Fixed cells were digested with 0.1 mg/ml zymolyase-100T (US Biological) and 1/10 volume of glusulase (PerkinElmer) at 30C for 15 min, washed once, and resuspended in 1.2 M sorbitol in 0.12 M K2HPO4/0.033 M citric acid, pH 5.9. Primary antibodies were anti-HA monoclonal antibody (HA.11; 1:500) and anti-tubulin (Abcam; 1:500). Secondary antibodies were: anti-mouse Cy3 (for HA) and anti-rat FITC (for tubulin). Cells were resuspended in DAPI (1 mg/ml). Imaging was performed at 25C using a Leica DM6000 microscope equipped with a 100x/1.40 NA oil immersion objective lens, A4, L5, and TX2 filters, and a digital CCD camera (DFC350, Leica). Pictures were processed with LAS AF (leica) and ImageJ software.

Biological Sources
NCBI Organism Classification
Saccharomyces cerevisiae
Cell Line
Cellular Component
spindle pole body
spindle pole body
tubulin complex
Mauricio Valerio-Santiago
Fernando Monje-Casas
J Cell Biol. (2011) 192: 599-614.
Digital Object Identifier (DOI)
Archival Resource Key (ARK)
Grouping This image is part of a group.
Sample Preparation
formaldehyde fixed tissue
Relation To Intact Cell
whole mounted tissue
Spatial Axis Image Size Pixel Size
X 187px 0.0642µm
Y 187px 0.0642µm