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BioFrontiers Advanced Light Microscopy Core

Welcome to the BioFrontiers Advanced Light Microscopy Core, a user facility designed to advance biological discovery through quantitative microscopy techniques.

Arbitrary pixel sizes in laser scanning microscopes permit low-magnification (40x), yet high-resolution (approximately 200 nm), imaging.

The low signal levels that result from biologically important processes, as well as the spatio-temporal dynamic nature of these processes (3D; millisecond timescales; longitudinal observation), necessitates the use of versatile, reconfigurable, high-speed, and high-sensitivity imaging technologies.  Multiple imaging technologies are housed within the BioFrontiers Advanced Light Microscopy Core that can meet these rigorous demands, ranging from conventional widefield to state-of-the-art Super Resolution/localization microscopies.

Current demands in biological research require the capability to observe cellular and sub-cellular events and phenomena both punctually as well as over time.  The BioFrontiers Advanced Light Microscopy Core is capable to meet these imaging needs. For example, the Nikon A1R resonant scanning confocal system is capable of imaging high-resolution z-stacks, using up to 7 different laser lines suitable for a large suite of fluorophores. Further, the A1R can simultaneously detect 4 different emission filters allowing for rapid imaging of multiple fluorophores within living samples. Using the resonant scanner, images can be acquired at up to 420 frames per second, easily capturing the fastest biological events (neurotransmission, calcium signaling, etc.).  Additionally, it provides the capability of ultra-sensitive total internal reflection fluorescence (TIRF) illumination with single molecule imaging and time-resolution on the order of 10's of milliseconds.

Complementary to the punctual and rapid observation of cellular and sub-cellular events and phenomena, the continuous long-term observation of multiple cell populations is also possible due to the implementation of high-end autofocusing technology along with full environmental control (temperature, humidity, oxygen and carbon dioxide).  This is available on multiple imaging systems within the facility.

Discovery based techniques, including high-content imaging of pharmacological perturbations or RNAi based screens, can also be carried out in 96 or 384-well plates. With experience in automation of image acquisition, as well as automated image analysis, vast improvements in productivity can be accomplished for many of the microscopes within the JSCBB microscopy core.

 

The following techniques are available for use:

  • N-STORM Superresolution Imaging
  • Confocal Imaging with 4 Detectors and 6 Excitation Lasers
  • Widefield Fluorescence Imaging
  • Total Internal Reflection Fluorescence (TIRF) Microscopy
  • Fluorescence Resonance Energy Transfer (FRET)
  • Calcium Imaging
  • Long-Term Imaging with Temperature, Oxygen, Carbon Dioxide, and Humidity Control.
  • Sub-Diffraction Super-Resolution Imaging
  • Differential Interference Contrast Microscopy
  • Photoactivation
  • Fluorescence Recovery after Photobleaching
  • High-Content Screening
  • Volumetric Imaging

 

More detailed information regarding the microscopes available can be found within the User Resources section.