Selective Plane Illumination Microscopy (iSPIM/diSPIM)

There are three basic configurations of SPIM systems 

that we have built for various users.

Fixed Sheet systems: 

The light sheet is stationary, with only mechanical adjustment for the sheet position. 

The two objectives are manually adjusted to correctly focus on the sheet. 

The specimen is scanned through the sheet using the X and Z stages to generate volume images.

Advantages: 

Least expensive – not requiring either galvo scanners or a piezo objective positioner.

Disadvantages: 

difficulty in correctly overlapping the objective focal planes with fixed positioners 

and the relatively slower stage scanning at odd angles.

Standard single-sided system: 

Light sheet from one side, emission objective on the other. 

The light sheet can be scanned using galvos to sweep across the sample volume. 

There is an emission objective piezo so the viewing objective can be positioned 

to follow the light sheet as it is scanned through the sample.

Advantages: 

Rapid Scanning, straight-forward setup.

Disadvantages: 

Better XY resolution than Z resolution.

Double-side system: 

Light sheet excitation and emission on each side. 

Both sides have a piezo-objective positioner. 

You need light sheets on each side as well. 

Advantages: 

With proper post processing, XY & Z resolutions are all very good 

– yielding a combination of speed and resolution that is unsurpassed for live cell imaging.

Disadvantages: 

Complicated and expensive.

Published Papers on the iSPIM/diSPIM System

Wu at al. Nature Biotechnoloty (2013)

Wu et al. PNAS (2011)

Kumar et al. Nature (2014)

Recent Webinars

Imaging Biology at High Spatiotemporal Resolution