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Dualview Inverted Selective Plane Illumination Microscope (DISPIM)

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Harshad Vishwasrao , Ph.D.
Photo of Harsh Vishwasrao
Acting Director
 Labs at NIBIB

The DISPIM is a dual light sheet microscope that acquires two orthogonal views of a sample. These two views can then be fused and joint deconvolved to generate a single volume with high isotropic resolution. Like other light sheet microscopes, the DISPIM provides rapid volume imaging with a low photodose and is therefore ideally suited for long term or high repetition rate imaging in samples where phototoxicity/photobleaching can be a problem. Unlike conventional light sheet microscopes, the joint deconvolution process provides good resolution in all directions.

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Image showing instrumentation of single molecule imaging microscopy
Figure 1. Spatial resolution: 330nm isotropic. Temporal resolution: 200Hz framerate, 1-5Hz volumetric. Excitation wavelengths: 405nm, 488nm, 561nm, 639nm. DISPIM is ideal for imaging experiments in which photobleaching or phototoxicity is a critical issue. This is the fastest microscope currently available at AIM.

 

Dual view example of DISPIM microsope
Figure 2. 40X 0.8NA, dual views provide 330nm isotropic resolution with low photobleaching/phototoxicity. Ideal for prolonged imaging of live samples.

 

Dual view image of cleared mouse heart
3. 17X 0.4NA, dual views provide lower resolution (~1um lateral) with a larger FOV (800x800um). Image plane can be scanned rapidly through a large sample of cleared tissue (up to 1cm x 1cm x 5mm). Refractive index: 1.33-1.56. Cleared mouse heart (courtesy Will Kowalski, NHLBI). 1.6mm x 2.6mm x 780um. Yellow: Anti-Tuj1-Alexa488 (pan-neuronal), red: Anti-SM22alpha-Alexa594 (smooth muscle) Acquisition time: 608s for 3 colors. iDISCO.

 

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