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Laser Scanning Confocal Microscope (LSCM)

A laser scanning confocal microscope is similar to the light microscopes that frequent most research labs, but utilizes laser light of differing wavelengths for enhanced imaging capabilities. The main advantage to this scope is the ability to remove all out-of-focus light, which aids in the creation of 3-D sample reconstructions. This allows researchers to accurately determine structure depth and placement.

LSCM Equipment 

The lab has a Leica SP2 LSCM.  The scan head can be mounted on either a semi-automated up-right Leica DMRXE7 light microscope, or on a fully automated DMIRE2 inverted light microscope.  Both microscopes are equipped with mercury burners, and full DIC optics.  Objectives are available ranging from 5X through 100X, including 20x and 40x dipping lenses to be used on the upright, and 40x and 63x objectives with correction collars to allow you to see through culture dishes and plates on the inverted. 

Excitation wave lengths
458nm, 476nm, 488nm, 514nm, 561nm, and 633nm.  

Emission detection
Precise emission spectra can be selected as needed using the Acousto-Optic Tunable Filter (AOTF).  The selected spectra range is then collected by one of three Photomultiplier Tubes (PMT), before the image is generated on the monitor, using the assigned Look-up table (LUT)  (a look up table converts the raw signal into a color code, so you get something that looks familiar, instead of a table full of numbers).

Optical sectioning
The standard stage can move at 0.2 micro increments; if needed a galvo stage can be mounted to support z-spacing below 0.2 micro increments.  

Software
The Leica software comes with a suite of image analysis tools such as image projection and movie making tools, dye finders, and separation tools.

LSCM Preparation Techniques

Currently, the following techniques are used for research purposes:

  • FISH (fluorescent in situ hybridization)
  • Co-localization
  • Epitope Tagging (typically using antibodies)
  • Auto-fluorescence for high resolution plant tissue reconstruction
  • Fluorescent Proteins such as GFP

Contact the Biological Imaging Lab

Mailing Address
Department of Biology
Biological Imaging Lab
Gottwald Science Center
28 Westhampton Way
University of Richmond, VA 23173

Office phone: (804) 484-1541
Lab phone: (804) 289-8775

Questions about the Biological Imaging Lab? Contact:
Christine Davis (804) 484-1541