STORM Super Resolution

Overview:  STORM and PALM are localization super resolution methods that provide up to a 10-fold improvement in resolution compared to conventional fluorescence microscopy.  These approaches temporally separate the emission of individual fluorophores in a sample with specialized imaging conditions.  “Localizations” are accumulated over time to build the final image.  Each molecules’ recorded fluorescence distribution is fit to a Gaussian function in post-processing to identify its position. The aggregated coordinates are displayed as a reconstruction.

This is an add-on modality to the Andor Spinning Disk/Widefield/TIRF system that can provide down to 20 nm lateral resolution in one or two channels.  There are a number of sample considerations for STORM/PALM imaging.  Please consult our sample prep guide.  The system is located in the Biology and Physics Building (BPB), room G05D.  Click here for rate information.

Super Resolution
Alexa 647 labeled microtubules imaged on the UConn STORM system. High laser power and STORM imaging buffer combine to induce photoswitching (blinking) of the dye molecules. Thousands of frames are acquired at high speed over several minutes.  Click on the image to see the movie.

 

STORM Modality Specifications:

Microscope: 

  • Inverted Stand Nikon Eclipse TiE
  • ASI Motorized x-y stage
  • ASI piezo z stage insert

Lasers: 

  • 405 nm photoactivation/conversion laser
  • 561 nm imaging laser
  • 633 nm imaging laser

Objective Lenses:

  • 100X/1.49 Plan Apo oil, DIC

Detector:

  • Andor iXon DU-897 back-illuminated frame transfer EMCCD

Software:

  • Micro-Manager for acquisition
  • ThunderSTORM ImageJ plugin for analysis

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