Housing for SiN Chip Vitrification – Team 2

Overview

In sample preparation of Cryogenic Electron Microscopy (Cryo-EM), blotting is tedious and problematic because it removes excess moisture from sample slides without consistency. Dr. Cameron Varano aims to design a new vitrification process to eliminate blotting.

Objectives

To design and construct a humidification system for a new Cryo-EM vitrification system that is able to reach 90% humidity in less than 20 minutes.

Approach

Overall Approach:
• We gathered our customer needs by scheduling weekly meetings with our sponsor. We asked lots of questions on the humidity we had to achieve. We even received hands-on training for the vitrification process.
• We created a screening matrix to identify the most important criteria as safety, functionality, durability, simplicity, and compatibility.
• We made a list of specifications from the screening matrix and determined that existing humidifiers would be more effective than any humidifiers we could build ourselves.
• We then searched online for solutions for regulating humidifiers and reviewed the specs of multiple options.
A prototype was created using a reptilian humidifier, hosing, and a humidity controller. The initial test determined if the prototype could maintain humidity in a closed environment. The protype successfully reached the target humidity in 15-20 minutes. Additional testing was planned to examine and prevent contamination inside the humidifier using biological assay testing methods. However, due to the COVID-19 situation, testing could not be finished.

Outcomes

As the goal of this project was to develop an entirely new and more accurate vitrification system, saving cost was not a primary concern of the sponsor. However, the team spent $122.19 in total, coming $877.81 under the allotted $1,000 budget. While humidification time was not reduced in this project, the prototype achieved target humidity within the allotted 20-minute time span, making it equally effective in comparison to the original humidification system. The prototype is extremely simple to operate, requiring the user to simply fill the humidifier, turn it on, and set the controller to 90%. This contributes to the overall enhanced simplicity of this vitrification system in comparison to the original system that contained inconsistent and costly blotting steps. The ultimate outcome of this project is a safe, functional, simple, and compatible humidification prototype that will allow Dr. Varano to proceed to the next steps in the development of her novel vitrification system.