Project 1B: Developing a New Voltage Reporter
Table of Contents
Introduction #
The key purpose of Project 1B is to develop a voltage reporting protocol that is sensitive enough for usage with non-excitable cells and can be used during cell reprogramming. We will determine through this whether a voltage reporting dye or a genetically encoded voltage indicator is more suitable for our purposes. The goal is for a reporting technique with a less than 5mV margin for error and high temporal resolution.
GEVIs vs Dyes #
| Benchmark | GEVIs | Voltage-Sensitive Dyes |
|---|---|---|
| Sensitivity | High sensitivity to voltage changes | Moderate to high sensitivity, varies by dye |
| Temporal Resolution | High temporal resolution | High, but can be limited by dye kinetics |
| Specificity | High, can be targeted to specific cell types | Lower, stains all cell membranes |
| Invasiveness | Non-invasive genetic encoding | Invasive, requires dye loading |
| Phototoxicity | Lower phototoxicity | Can be phototoxic, especially under prolonged exposure |
| Signal-to-Noise Ratio | High, due to specific expression | Lower, due to background staining |
| Stability | Stable expression in cells | Can be unstable, prone to photobleaching |
| Ease of Use | Requires genetic manipulation | Easier to apply, just add dye |
| Multiplexing | Limited by available spectral variants | More flexibility with different dyes |
| Cost | Higher initial setup cost | Lower initial cost, but recurring expenses for dyes |
At the time of writing this, rEstus1 is our leading GEVI candidate and Di-4-ANEPPS is our leading dye candidate.
Rühl P, Nair AG, Gawande N, Dehiwalage SNCW, Münster L, Schönherr R, Heinemann SH. An Ultrasensitive Genetically Encoded Voltage Indicator Uncovers the Electrical Activity of Non-Excitable Cells. Adv Sci (Weinh). 2024 Mar 25:e2307938. doi: 10.1002/advs.202307938. Epub ahead of print. PMID: 38526185. ↩︎