Live Animal Imaging Confocal Activates Vascular Clotting

On a recent project I had an opportunity to collaborate with some great manufacturers and vendors, to build a custom microscope for Dr. Chia-Yi (Alex) Kuan, at the University of Virginia School of Medicine. This instrument is quite unique, and required a team of manufacturers to pull it off.

Completed system ready for use.

Dr. Kuan required an upright confocal microscope, which could perform photoactivation, widefield and confocal fluorescent excitation, all while running on an extended focus throw of 2″ range. The system required several custom pieces of hardware and software, which made things extra *fun*.

This image was taken from a live specimen, showing cranial vasculature in vivo.

 

 

 

 

 

 

Team Contributors

Visual Dynamix LLC, headed by Gary Tockman, was the lead system integrator on the project as well as the primary service provider for the system. Gary led the team on the build and did an excellent job defining input specifications and performance criteria, as well as managing the crazy logistics of building a system from so many different vendors.

Using the same device for slides and for live specimens required some work on the focuser throw. Sutter Instruments provided a great solution via the BOB microscope. Chris Ballard spent a great deal of effort working out a long travel Z motor relying on the MPC control system for XYZ axis.

89 North provided the photostimulation system, confocal, and LDI laser excitation. Scott Phillips and Simona Stelea both spent several days on site working on the system to make sure everything was dialed in properly.

Scott Phillips works on cable connections

All of the devices on the system needed control via Metamorph. I worked with David Biggs over at KB Imaging to handle this part, and was very glad I chose his work. David has extensive experience with driver development, and due to the proprietary nature of the firmware on this instrument, his experience saved the day on dialing in the driver for speed and reliability.

Finally, I took on the system I/O side for lasers and triggering, as well as building the imaging PC and configuring Metamorph. Because the latest Triggerscope 3 model needed to drive the lasers,I also managed to work with David to create a Metamorph driver, so if you happen to run MetaMorph and want to control the triggerscope, please get in touch.

Gary inspecting the filters on the XLight

 

 

 

 

 

Systems like this usually require a day or two for the install, but in our case, most of the install team spent a week on site, to be certain that the details were ironed out and to get some training on the system in for the end users. In our situation this was a good call, as we found the inevitable last minute changes which needed to be addressed. Having the team together made addressing these things straightforward.

Results

A bit after leaving, the client sent over a beautiful video showing experimental data which includes the visualization of vasculature, with a clot stimulated by the laser exciter, and a resulting timelapse of that clot forming.

 

Training for end users on a live specimen

 

It’s great to be able to participate with clients, manufacturers and engineers who can pull builds like this together, and better still to watch this important and exciting research move forward. I’m even happier I get to share this build!

-Austin