Scripps Research Logo

Nikon Total Internal Reflection Fluorescence

Nikon Total Internal Reflection Fluorescence (TIRF) Super-Resolution Microscope (Room MBB34E)

The Nikon TIRF/STORM super resolution system is capable of fast live cell imaging in TIRF mode as well as super resolution imaging of fixed as well as live samples.

Hourly Rate for Usage: TSRI trained user hourly rates $25.00 from 9AM-5PM Monday-Friday,  $12 from 6-9PM and during weekends. $50 flat rate overnight.
Contact kpestonj@scripps.edu for training.

TIRF microscopy is designed to visualize fluorescent probes in a thin plane within ~100 nm of the coverslip. This provides high sensitivity due to the absence of background fluorescence from other specimen planes. The greatly enhanced signal to noise makes TIRF microscopy particularly useful to investigating cytoskeletal dynamics and vesicle trafficking. Stochastic Optical Reconstruction Microscopy (STORM) can be used to image cellular structures at 10-fold greater resolution than standard fluorescence microscopy (20 nm in the X-Y plane, and 50 nm in the Z-axis). This allows more precise imaging of supramolecular structures such as membrane receptor assemblies, viral capsids, mitochondrial sub-components, vesicular trafficking intermediates, focal adhesions, nuclear substructures, and cytoskeletal filaments, which have dimensions in the range of 10s of nm.

 

The Nikon TIRF/STORM system is equipped with the fast Hamamatsu Flash 4 camera and the latest version of the Nikon software (Elements) that enables acquisition of images at extremely rapid rates (a few milliseconds for the TIRF and a few hundred milliseconds per frame for STORM) allowing real-time live-cell imaging in both TIRF and STORM mode. The system is equipped with a high power LUNV laser launch with 4 lasers (405/488/561/647 nm). The high power lasers allow the use of most of the standard fluorescent tags that are used for conventional microscopy (eg. Alexa 488/568/647). Live super-resolution imaging is possible in 2 channels simultaneously. It is equipped with the following infinity corrected optics:


• 10X 0.3 NA PlanFluor Ph1 DLL
• 60X 1.45 NA Oil Plan Apo TIRF DICH
• 100X 1.49 NA Oil HP Apo TIRF DIC N2