Light-Sheet Microscope

 

Slicing tissue samples to image them can destroy or alter delicate structures. Most modern microscopes can image a maximum of a few millimeters, so slicing is necessary. Light-sheet fluorescent microscopy can image whole organs very quickly, allowing for visualization of structures in delicate organs like the brain or cochlea that would otherwise be impossible to view.

The Greenbaum Lab

My Work

Undergraduate Researcher

I worked to complete one of the lab’s primary projects: creating a light-sheet microscope to image samples rendered transparent by other lab members. I was responsible for the construction of the microscope and corresponding software. The first step was adapting old schematics to our needs. I then researched and purchased the necessary optical components, and set up these components on our optical table. I was responsible for reworking an old Matlab script and custom function to be used with the new setup. Once new graduate students were hired, I trained them on the hardware and software of the microscope. After the microscope was constructed, I was given the project of aligning the lasers, which is a lengthy and delicate process. This microscope is critical to the current function of the lab, and will be used in nearly all future projects.

Administrative Assistant

I was Dr. Alon Greenbaum’s first employee. I worked initially within the role of administrative assistant to set up his new lab. I singlehandedly placed orders for every piece of equipment and reagent that was required for the first two years of the lab’s operation, totaling well in excess of $100k spent. This required maintaining accurate records of all orders and being the primary point of contact between the lab and vendors. I also worked with various companies to create an assortment of custom components for the lab or to problem solve issues with equipment. As the lab hired more employees, I was responsible for training them on the lab’s safety and waste disposal protocols and ensuring these protocols met the university’s standards.

  • Team

    This is the first group of students in the Greenbaum Lab, summer 2019.

  • Microscope in Progress

    Shown is the microscope setup. The table is a pneumatic optical table, which uses air pressure to eliminate vibrations or other extraneous motion. The optical elements are attached to the threaded holes on the table and other equipment is on the shelves above.

  • Hamamatsu ORCA-Flash4.0

    Microscope Camera

    Here is the camera used to image samples, a Hamamatsu ORCA-Flash4.0. Also pictured is the ASI tube lens used to focus the image.

  • Imaging Chamber

    We had a custom aluminum and glass imaging chamber made for this project, which was then filled with our imaging medium (dibenzyl-ether). Visible in this picture is the first light sheet we saw from the microscope (green laser line).

  • Two pinholes used to align the lasers

    Microscope Alignment

    Here’s an example of the process used to align the lasers. The laser is shined through two pinholes of the same height onto a light intensity reader. If the laser passes through both pinholes without losing much intensity, it is parallel to the table.

  • Galvanometer Mirror

    Galvanometer Mirror

    These mirrors are used to direct the path of the lasers, allowing the samples to be scanned. They move in tiny amounts proportional to the input current.