Dual-View Inverted Selective Plane Illumination Microscopy: diSPIM
The benefits of Selective Plane Illumination Microscopy, SPIM, have been acknowledged for some time now. These include reduced photo-bleaching and high acquisition speeds.
In SPIM, often referred to as lightsheet microscopy, a sheet of light is used to define a focal volume. This “lightsheet’ selectively scans through a sample with the fluorescence being detected perpendicular to the sheet of light.
Despite the benefits of SPIM over existing techniques, limitations exist which limit the applicability of SPIM. These include; sample preparation and fixing. That is, the sample needs to be mounted in a specific holder at a specific orientation which limits the technologies ability to say live-cell imaging samples on a glass coverslip, and the axial resolution, as with most imaging techniques, is compromised compared to lateral resolution.
A recent development by Dr. Hari Shroff of the NIH brings a novel development to SPIM to address the above shortcomings of the SPIM technology.
Dr. Shroff’s group has developed so-called Dual-View inverted Selective Plane Illumination Microscopy, or diSPIM. View the video on You Tube.
In diSPIM, a two objective lenses are used such that one lens generates the so-called lightsheet whilst a second lens positioned at 90° collects the fluorescence. A galvanometer is used to scan the lightsheet through the sample which can be mounted on a glass coverslip on an inverted microscope.
The limitations of axial resolution are addressed by reversing the architecture. That is, switching the objective lenses used for generating the lightsheet and fluorescence collection.
Applied Scientific Instrumentation, ASI, offer a range of systems solutions for both iSPIM (inverted selective plane illumination microscopy) and diSPIM.
Coherent Scientific is co-sponsoring a diSPIM workshop at the upcoming ACMM meeting in Melbourne. At that workshop, Dr. Hari Shroff will take participants through the benefits of iSPIM and diSPIM and specifically demonstrate the building a diSPIM microscope.
Ben Hibbs, of Coherent Scientific will be attending the workshop and we encourage you to speak with Ben regarding applications where diSPIM maybe beneficial in your laboratory.