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

CIL:11991*  Cite 
Description

TIRF time-lapse imaging of EB1-GFP (green) and mCherry-tubulin (red). EB1-GFP is seen on microtubule tips that grow to the cortex but does not accumulate on stable microtubule bundles at the equator, in contrast to kinesin-6 . Corresponds to video 8 from J Cell Bio 186:727-738, 2009. Total internal reflection microscopy was performed using a microscope (Perfect-focus TE2000; Nikon) with a 100×, 1.45 NA objective (Nikon) and illumination from either a 488-nm argon laser (100 mW) or a 491-nm solid-state laser (100 mW) and a 561-nm solid-state laser (50 mW). For dual-color TIRF microscopy, we used a triplepass dichroic filter (z491/561/633rpc) and changed the emission filter (ET525/50 or ET595/50; both from Chroma Technology Corp.) with a filter wheel placed before the camera. In some cases, an excitation notch filter was used in the filter cube (NF01-405/488/561/635; Semrock, Inc.). Images were typically captured every 2-3 s with a 50-200 ms exposure with an EM charge-coupled device camera (iXon; Andor Technology). The microscope was controlled and images were acquired using open source MicroManager software (http://www.micro-manager.org).

Biological Sources
NCBI Organism Classification
Drosophila melanogaster
Cell Type
epithelial cell
Cell Line
S2
Cellular Component
EB1
microtubule
Biological Context
Biological Process
microtubule polymerization
Attribution
Names
Ronald D. Vale
James A. Spudich
Eric R. Griffis
Published
J Cell Bio 186:727-738, 2009
Pubmed
19720876
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL11991
Archival Resource Key (ARK)
ark:/b7295/w9cil11991
Grouping This image is part of a group.
Imaging
Image Type
recorded image
Image Mode
TIRF
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
microtubule polymerization
mCherryFP
Processing History
unprocessed raw data
color combine
Data Qualifiers
raw, unprocessed data
Sample Preparation
Methods
living tissue
Relation To Intact Cell
dispersed cells in vitro
Dimensions
Spatial Axis Image Size Pixel Size
X 360px ——
Y 322px ——
Channel Wavelength
1 488, 561nm
Time 8 seconds


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