Longer description of your proposed project

Single-cell imaging allows researchers the unprecedented ability to study biological processes that occur over time spans from milliseconds to months. These processes include neural circuit activity, physiological changes within cells, signaling between cells, development, stem cell differentiation, cell motility, immune system activation, metastasis of cancer cells, development of tumors, drug kinetics, and more. Being able to track individual cells in a single field of view over many days has dramatically accelerated discovery and enabled researchers to look at the heterogenous physiology and changes in single cells (and even subcellular compartments), rather than population trends. Unfortunately, most labs, even relatively well-funded ones, cannot afford to use single-cell imaging.

Commercial systems start in the hundreds of thousands of dollars, unaffordable for nearly all labs, and the manufacturers have no competition and no incentive to decrease the cost. Our lab (rajasethupathylab.org) has developed a system for imaging using bundles of fiber optics that can accomplish single-cell imaging in three areas for about $30,000. The goal of this project is to develop a system using off-the-shelf components that will allow researchers without specific expertise in optics or single-cell imaging to build and run this system for under $10,000. The plans will be made available for free online. We have no plans to commercialize this – our goal is to make it so that labs anywhere around the world and without a lot of money to throw around can harness the power of single-cell imaging.

Our system is better than other approaches. Some use large and expensive microscopes that require a research animal to be head-fixed, which is stressful for the animal, limits the imaging time, and makes most natural behaviors impossible. Miniaturized microscopes are heavy and bulky, interfering with an animal’s movement and limiting the sites you can record to one or two, and are permanently attached to an animal, meaning the cost of running animals quickly rises. Our approach allows for recording from three or more areas, and even tiny mice can move around easily and comfortably, executing all of their normal behaviors.

Moreover, this approach improves animal welfare by increasing comfort, improving validity of results, and decreasing the number of animals needed. Not only will it allow for multiple regions to be visualized at once, but it replaces techniques that rely on histology. Instead of sacrificing and imaging tissues from animals at each time point, biological processes can be directly imaged without harm to the animal. Using this technique, animals remain comfortable and healthy for years.

Describe why you think you're qualified to work on this

I am a neuroscientist with multiple successful projects and years of experience in circuit neuroscience. I work as a postdoctoral fellow in the Laboratory of Neural Dynamics and Cognition at Rockefeller University. I am an expert in all techniques necessary for this project. My current and previous labs are known for technological innovations in neuroscience, including the fiber bundle technique we intend to make more accessible here. We enjoy access to a great machine shop and maker space, allowing us to prototype cheaply; however, our final design will be available to labs without these resources.

Other ways I can learn about you

My Twitter handle is @aluskin. My lab website is rajasethupathylab.com. I have two first-author papers, one in Science Advances (https://www.science.org/doi/10.1126/sciadv.abd3666) and one under revision at another major journal (https://www.biorxiv.org/content/10.1101/2022.06.30.498327v2).

How much money do you need?

$25,000 – the money will be spent testing different components, with the goal of landing on a configuration that will cost $10,000 or less.

Links to any supporting documents or information

See https://www.cell.com/cell/fulltext/S0092-8674(23)00167-8 for a description of the fiber bundle technique; see https://www.rajasethupathylab.com/resource-fiberbundletechnology for a guide to using the fiber bundle technique we have already developed. We are developing a low-cost version of this system.

Estimate your probability of succeeding if you get the amount of money you asked for

99% (ie as long as I don’t get pushed in front of a subway train) - building a working rig. 90% - building a working rig that will cost the final user under $20,000. 80% - building a working rig that will cost the final user under $10,000.