stun : High Throughput Neuron Injury Device
Neurotrauma and Repair Lab at Columbia University
Summer 2013, Summer 2014
I spent two summers as a research assistant in Dr. Barclay Morrison's Neurotrauma and Repair Laboratory at Columbia University. I was working for Dr. John Finan, a post-doctorate student at the time, designing a semiautomatic neuron injury method for quick, precise, repeatable, cell injuries in a 96-well plate for pharmaceutical testing in high throughput screening machines.
INJURY DEVICE PROTOTYPE
WELL PLATE CLAMP WITH PTFE PISTONS
PISTON MECHANISM SECTION
Dr. Morrison had invented a device that stretched neurons grown on a silicone membrane over a post to simulate blunt traumatic brain injury. While the device had been successfully used for years and yielded valuable data, it was labor intensive, high cost, and low throughput. My project was to prototype an injury device compatible with high-throughput screening robots; the ultimate research goal being to explore novel treatments for traumatic brain injury (TBI) by testing all known bioactive compounds (~100k) on injured neurons. Neurons do not show signs of TBI until ~24h after injury (part of why it can be so difficult to detect concussions), and this presents a possible window for treatment.
DR. MORRISON'S DEVICE
INJURY MECHANISM
To be compatible with high-throughput screening robots, the cells had to be cultured and injured in standardized multi-well plates. I invented a flexible bottomed 96-well plate for cell culture. I decided to make the well plates out of commercial polystyrene bottomless well plates and thin silicone membranes to keep testing costs down and ensure that the well plate manufacturing procedure could be scaled up. After weeks spent researching chemical bonding techniques, I figured out how to bond the two inert materials without glue so the final product would be durable and safe for cell culture. I documented the procedure, which involves activating the polystyrene in a plasma cleaner and applying the aminosilane APTES to form a covalent bond with the silicone membrane.
PLASMA GLOW DURING TREATMENT
UNDERSIDE OF A FLEXIBLE-BOTTOMED 96-WELL PLATE
The injury device replicated blunt traumatic brain injury by using a 1.4kN peak-force, 20A linear actuator to slam the well plates down on Teflon posts, stretching the silicone membrane and the neurons growing on it to induce strains up to 200% in 50ms. I chose to actuate the well plate rather than the posts so the membranes would remain in the same plane throughout the stretch, enabling accurate strain measurement with a high speed camera (frame from a characterization video above — wells 1-3 are not stretched, wells 4-6 are mid-stretch).
BEFORE INJURY
MID-INJURY
Dr. Finan now leads a laboratory at NorthShore University Health System that uses this high-throughput injury device as a fundamental part of its in-vitro neurotrauma model.