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CIL:11924*  Cite 

Comparison of actin speckle microscopy with phalloidin staining. Primary cultures of newt lung epithelial cells were microinjected with X-rhodamine actin. Time-lapse FSM was performed on a spinning disk confocal microscope system. Light from a 50-mW Krypton-Argon ion laser (Melles Griot, OmniChrome) was delivered by a single-mode fiber optic (Point Source) to a Yokogawa spinning disk confocal scan-head (Ultra-View; PerkinElmer) on an inverted microscope (TE300 Quantum; Nikon). Excitation wavelength was selected by a filter-wheel apparatus (Sutter Instruments Co.) containing excitation filters for 488 and 568 nm (Chroma). Emission was selected by multiple bandpass dichromatic mirror and emission filter (Chroma). Images were collected by a 100x 1.4NA Plan-Apo DIC objective lens (Nikon) and captured with an Orca 2 camera (Hamamatsu). Images were collected in the ventral focal plane at 10s intervals. Following live cell imaging, cells were fixed with paraformaldehyde, permeabilized and stained with Alexa 488 phalloidin. Images were processed as follows: (1) background subtraction; (2) 3 x3 low pass filter; (3) unsharp mask filter. Video corresponds to Fig 1 and video 1 in J Cell Biol, 158:31-37, 2002.

Biological Sources
NCBI Organism Classification
Taricha granulosa
Cell Type
epithelial cell of lung
Cellular Component
actin cytoskeleton
Wendy C. Salmon
Michael C. Adams
Clare M. Waterman-Storer
J Cell Biol, 158:31-37, 2002
Digital Object Identifier (DOI)
Archival Resource Key (ARK)
Grouping This image is part of a group.
Image Type
recorded image
Image Mode
spinning disk confocal microscopy
fluorescent speckle microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
Alexa Fluor 488
Processing History
background subtraction, 3X3 low pass filter, unsharp mask
Data Qualifiers
processed data
suitable for spatial measurements
Sample Preparation
living tissue
formaldehyde fixed tissue
Relation To Intact Cell
dispersed cells in vitro
Spatial Axis Image Size Pixel Size
X 371px 0.119µm
Y 337px 0.119µm
Time 10 seconds 8.3 min