Bright-field illumination

Michael Redd from the Cell Imaging Core Facility at the University of Utah put together an Arduino-based solution to get proper bright-field illumination in an OpenSPIM setup: Brightfield.

Shutter

Instructions to make an Arduino-controlled shutter compatible with uManager: paper by Todd P. Meyrath and tutorial on the uManager page.

Stepper Motor Upgrade

If you would like to swap out one of the Picard stepper motors you can do so with a bit of effort.

OpenSPIM for large samples

Field of view

The original OpenSPIM uses a 2X expander to enlarge the laser beam to 2 mm. For larger samples a larger field of view can be preferable. This means that you will need a larger beam to create a taller light sheet. Change the second beam expander lens to f=75 mm or 100 mm and increase the distance between lenses accordingly.

Objectives

Immersion objectives with long working distance are hard to find. However, at low NA and moderate magnification, a dry objective will also work; the higher NA, the more spherical aberrations will be present. Keep in mind that when using a dry objective in immersion, the WD will increase (approximately by a factor equal to the RI of the new medium, eg. 17 mm in air will increase to 22.6 mm in water).

OpenSPIM for brains setup

Click here for a table that lists the parts used by the OpenSPIM for brains setup built by Monika Pawłowska (Nencki Institute of Experimental Biology, Warsaw).

X-OpenSPIM example equipped with two laser lines and a cooling chamber

A detailed description of how an OpenSPIM can be upgraded to contain a second illumination axis with two cameras and multiple laser lines under the control of an Arduino board. The system also features a cooling chamber.