Cell Imaging...Laser Scanning Confocal  
Contact info:
Chris Rodesch
crodesch@cores.utah.edu
Phone: 587-7964
Fax: 585-6364		 Back to Cell Imaging Home

  • Olympus Fluoview™ series confocal microscope
  • Argon and krypton lasers providing excitation energy at 488, 568, and 648 nm for use with single-, double-, and triple-labeled specimens
  • simultaneous acquisition of fluorescence and transmitted light (brightfield or DIC) images
  • water immersion objective lenses (40x and 60x) for use with aqueous samples
  • 100x, N.A. 1.35 oil immersion lens for high resolution work
  • software enables 3-D rendering, thresholding and binary operations, image filtering, arithmetic operations (ratioing), quantitative analysis
  • a freeware distribution (ImageJ, a Java-based version of NIH Image) is available with plug-ins for use with Fluoview-formatted image files

Examples:
Mitochondrial Staining in Mouse Muscle Cells:		  


Morphology and distribution of mitochondria were analyzed in C2C12 mouse myoblasts. Cultures were incubated in 25 nM Mitotracker Orange (Molecular Probes) in medium for 20 m. The image shows the projection of a z-series comprising 16 slices acquired at 0.2 um intervals.

Stained cells and protocol courtesy Brandi Baker and Dr. J. Rob Singleton, Department of Neurology.

 
 

Features difficult to observe in the projection image can sometimes be more easily discerned after rendering the 3-dimensional volume through a rotational series.

Click the play button on the left, drag the frame slider, or click the scroll buttons on the right to view the animation.
QuickTime, which can be downloaded here, is required to view the rotational series.



Identification of apoptotic cells in colonic crypts:


An immunofluorescent study was used to examine the distribution of apototic cells in colonic crypts. Apoptotic cells are stained green with an Alexa 488-conjugated secondary antibody (Molecular Probes); total nuclei are stained red with propidium iodide.

Image and protocol courtesy Phil Gray, Department of Gastroenterology, and Dr. James Disario, Department of Internal Medicine.

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