ASI´s New Dual Inverted Selective Plane Illumination Microscopy (diSPIM)

  ASI has developed a new form of light sheet microscopy with our collaborators within the scientific community. The system utilizes two water emersion objectives mounted at 45 degree angles above the sample on an inverted microscope, a third objective also can be used to view the sample from below. The top objectives are mounted on piezo positioning devices so precise image stacks can be obtained. The system utilizes micro-mirrors to generate the light sheet, and the complete system can be mounted on to an existing inverted microscope, or supplied with ASI’s Rapid Automated Modular Microscope system (RAMM). Some of the benefits of the system include:

• Use conventional mounting /glass cover slips
• Generate 3D volumes with isotropic resolution (330 nm in all directions)
• Axial resolution is ~2x better than confocal- or spinning disk systems
• Achieve a ~7-10 fold reduction in photobleaching
• Acquisition rates up to 200 images per second or 2-5 volumes per second (roughly 10x faster than spinning-disk microscopes)
• Compared to Bessel beam plane illumination approaches, diSPIM offers equivalent (or better) axial resolution (~330 nm), 10-100x faster volumetric imaging rates (0.5-1 s instead of 10-100 s), and the ability to image over ~10x more time points (~1000 instead of ~100), due to the significantly lower illumination dose employed in our experiments and the lack of extraneous illumination outside the focal plane
• Microscope has multicolor capability, and has been tested successfully on cells cultured on cover slips, cells embedded in collagen gels, and nematode and zebrafish embryos.
Micrasys e.K.

Frank Pobig
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