Fluorescence microscope enabling time-lapse tracking of compact, multistable gene circuits for cellular regeneration
Abstract
We propose to purchase a fluorescence microscope that will be used to image cells undergoing programmed fate transitions over longer time scales (days). This instrument combines a large 25 mm field of view with a focus tracking system that accurately tracks locations relative to the cover slip surface, enabling high resolution time-lapse imaging with no drift over time. This high content imaging will permit the study of mammalian cell population dynamics and identification of gene expression kinetic parameters through detection of mRNA and protein species over time. Unlike plate-readers and flow cytometers, this fluorescence microscope can be used in conjunction with existing analysis pipelines to study single-cell traces. Additionally, this dynamic analysis will complement our existing steady state flow cytometry capabilities for a more complete understanding of how cell-fate transitions progress over time. The instrument will be utilized in multiple DOD-funded research projects in mammalian gene circuit design for robustness and composability. Specifically, we will use the system to build a proof-of-concept regeneration model capable of autonomously responding to injury. The dynamic analysis afforded by this setup will permit us to assess the robustness of the proposed circuit in response to time-dependent disturbances. If successful, the proposed research will accelerate the design-build-test cycle of mammalian synthetic biology through establishing design rules for compact, multi-gene systems controlling cell fate transitions. We expect the increased efficiency and throughput of this instrument will also enable us to develop new capabilities for visualizing transcriptional burst dynamics, leading to advances in model-guided gene circuit design.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Feb 05, 2025
- Source ID
- FA95502410052
Entities
People
- Kate Galloway
Organizations
- Air Force Office of Scientific Research
- Massachusetts Institute of Technology
- United States Air Force