Core Facilities

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Confocal Microscopy

The Confocal Core Facility is located on the second floor of the new Arthur C. Guyton Research Building in Room G272. The Confocal Core houses a Leica TCS-SP8 multiphoton confocal microscope. The system has three separate lasers for use with 3 standard visible length fluorescence dyes as well as transmitted light. The multiphoton includes a separate infrared laser.

Confocal Core Services

The SP8 is equipped with three individual lasers (488/561/633 nm) capable of imaging standard fluorescence dyes within the visible spectra including FITC/TRITC, CY2/CY3/CY5, green fluorescence protein variants (GFP/YFP/RFP/DsRed) and many different indicator dyes (i.e. Ca2+, pH, membrane potential, oxidative stress, etc). This system uses a light green laser (561) instead of the 546 green laser. The 561 laser can still be used with standard CY3 and TRITC dyes, but Alexa 555 dyes are ideal. The multiphoton capability is provided by a Coherent sapphire laser. This system includes an inverted microscope with a Luden environmental chamber for temperature control.

A researcher uses the SP8 confocal microscopeCapabilities

  • Epifluorescence provided by a halogen lamp (SP8)
  • Motorized stage
  • Standard confocal microscopy with simultaneous or sequential image collection
  • Line scan, Z-scan modes
  • High speed resonant scanner
  • High resolution imaging
  • Transmitted light detection
  • Tunable collection filter
  • Time lapse
  • Stage adaptable for slides, dishes up to 35mm,  25mm glass cover slip in an atto chamber 
  • Image enhancement
  • Quantitation, 3D reconstruct
  • High speed resonant scanner
  • High resolution image
  • Objectives: 63X glycerol, 1.37 NA, 40X oil 1.25 NA, 20X dry 0.7 NA, 10X dry
  • Super Z galvo stage
  • 6PMTS: 3 color imaging (includes 2 high sensitivity hybrid detector, standard PMT), 1 transmitted light PMT, 2 non-descanned light detectors
  • Inverted orientation permits fixed or live cell/tissue imaging.
  • FRET/FRAP (Fluorescence resonance excitation transfer/ Fluorescence recovery after photobleaching)
  • Quantitation and colocalization
  • 3D Reconstruction
  • Multiphoton imaging (deep tissue imaging, dark/dense tissue far red imaging)

Fees

  • Training: $100/hr
  • User-based imaging: $50/hr

The Confocal Microscopy Core is primarily an equipment core. Users new to confocal microscopy typically require 2-4 hours of training depending on level of microscopy experience. Experienced users typically require 1-2 hours of training.

Acknowledgement of Core Services:

This Core serves an invaluable resource for many users at UMMC and is supported by multiple NIH funded programs as well as the Department of Physiology & Biophysics. Your use of this core is greatly appreciated and we hope to continue to provide these services to support your research program. Sustainability of these core services is a major goal of the Physiology Department.

 As a user of these core services, you are expected to acknowledge the proper funding source that provides support for your use of this core. For every publication, press release or other document regarding above mentioned use of these core services, the appropriate NIH award, Cardiorenal and Metabolic Diseases Research Center P30GM149404 should be cited and acknowledged within PubMed Central (PMC).  Appropriate acknowledgement can be reported as follows: "Research reported in this publication was supported by the National Institute of General Medical Sciences of the National Institutes of Health under Award Number P30GM149404. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health."

However, if a manuscript includes one or more co-authors whose listed affiliation is with a foreign institution, and the publication acknowledges support from the NIH IDeA Program, including the COBRE award,investigators should contact the COBRE Administrative Core before manuscript submission. NIH has recently emphasized that publications acknowledging IDeA funding and including authors affiliated with foreign institutions may require prior review and approval by NIGMS, even when the research activities were conducted entirely in the United States. The COBRE Administrative Core will help determine whether additional documentation or consultation with NIGMS is needed before the manuscript is submitted or the award is cited.

For more information, Contact:

Heather DrummondHeather Drummond, PhD
Office: G258
(601) 984-1812