Dynamic live cell imaging at sub-zero temperatures
Lead Research Organisation:
NERC BRITISH ANTARCTIC SURVEY
Department Name: Science Programmes
Abstract
The challenge: The study of life's adaptation to extreme environments challenges our fundamental understanding of biological systems from molecular to whole organism levels. Proteins are key building blocks for all life on Earth with functions that are uniquely dependent on their 3-D folded state. Whilst much is known about constraints on how proteins operate at high temperatures, little knowledge exists about how biology operates at all scales of life in sub-zero conditions where proteins are less stable and oxidative damage is high. Almost 90% of the habitable biosphere is permanently below 5°C (i.e. deep sea and polar regions). Hence, we do not understand how a large proportion of global biodiversity functions at such low temperatures: A critical knowledge gap given the current climate crisis and impeding large-scale loss of the planet's colder regions and their endemic biodiversity.
Aims and interdisciplinarity: Cellular proteins are adapted to function in highly crowded solutions of macromolecules, which affect protein folding, diffusion, and interactions. Temperature plays a critical role in these processes. However, there are currently no tools available that image live cells at very low temperatures. We will use the most advanced methods to adapt current state-of-the-art microscopy, and for the first time, develop fully automated microscope technology optimised for the high-resolution optical imaging of live animal cells near 0°C. This will enable us to observe the behaviour of proteins in situ and gain a deeper understanding of the behaviour of proteins near 0°C within the complex environment of the living cell. The system will be used for studies of Antarctic fish cell cultures at 0°C, our cold-adapted model organism. In particular, we will study temperature effects and cell viscosity in the context of protein folding within the cell, using a fast-folding protein, Venus, introduced into the Antarctic fish cells at 0°C and use single molecule translation imaging, developed by us, to compare the time for protein folding with temperate systems. This highly interdisciplinary project is at the very intersection of biology, physics and chemistry and involves collaboration between world-leading researchers in cutting-edge microscopy, molecular cell biology, and polar marine biology.
Aims and interdisciplinarity: Cellular proteins are adapted to function in highly crowded solutions of macromolecules, which affect protein folding, diffusion, and interactions. Temperature plays a critical role in these processes. However, there are currently no tools available that image live cells at very low temperatures. We will use the most advanced methods to adapt current state-of-the-art microscopy, and for the first time, develop fully automated microscope technology optimised for the high-resolution optical imaging of live animal cells near 0°C. This will enable us to observe the behaviour of proteins in situ and gain a deeper understanding of the behaviour of proteins near 0°C within the complex environment of the living cell. The system will be used for studies of Antarctic fish cell cultures at 0°C, our cold-adapted model organism. In particular, we will study temperature effects and cell viscosity in the context of protein folding within the cell, using a fast-folding protein, Venus, introduced into the Antarctic fish cells at 0°C and use single molecule translation imaging, developed by us, to compare the time for protein folding with temperate systems. This highly interdisciplinary project is at the very intersection of biology, physics and chemistry and involves collaboration between world-leading researchers in cutting-edge microscopy, molecular cell biology, and polar marine biology.
