In 2015, in cooperation with the Institute of Instrumentation, CAS, we participated in the implementation of a system for "light sheet" microscopy based on the OpenSPIM project for the CEITEC Centre of Excellence.
Description of the equipment:
The excitation part of the system is equipped with 3 laser sources, which are brought into one optical path and guided into the sample chamber. The laser beam then passes through a set of lenses to form a rectangular light profile, which is finally incident through the lens on the sample in a chamber filled with immersion solution.
The sample chamber is tempered by a water circuit and is completely replaceable as a module. The sample is attached to a four-axis positioning system which allows movement in the X,Y,Z axes and further rotation along the Z axis.
The emission arm includes a motorized fluorescence filter holder as well as automated lens holders that allow for rapid image magnification changes. A high-resolution sCMOS camera is then used for data acquisition (imaging). The camera chip is thermoelectrically cooled to achieve low noise. The camera is mounted on a motorized travel, which solves the problem of easy refocusing when changing the magnification, or it can be removed by the user and used elsewhere.
The system is complete, encased in a single box mounted on an anti-vibration pad. A computer with free(open source) software for Micromanager acquisition and subsequent ImageJ data processing is included.
Components:
- Coherent OBIS lasers (488nm, 561nm, 647nm)
- Andor Neo 5.5MPx sCMOS camera
- Thorlabsoptics and optomechanics