Dr Siddharth Deshpande
Cees Dekker Group, Delft University of Technology
Dr. Deshpande’s research focuses on the use of synthetic biology approaches to study real-time dynamics of biomolecules.
While Cees Dekker’s group deals with diverse research projects, Dr. Deshpande’s work is focused on looking at biomolecular processes in vitro by applying bottom-up approaches to improve understanding of how living cells work. Specifically, the group studies key individual components of biological relevance within a confined space, such as liposomes or droplets, which are excellent scaffolds for creating synthetic cells. The size range of these containers is in the 5-50 micrometer range.
Dr. Deshpande mainly uses high-resolution time-lapse imaging to look at the dynamics of fluorescently labeled components, such as lipids, proteins, polymers, and DNA molecules, confined within microcontainers, to understand the self-organization and emerging behavior of these molecules.
Dr. Deshpande was previously using a front-illuminated sCMOS camera but wanted to improve upon the sensitivity to push the exposure time as low as possible whilst maintaining enough signal for imaging.
One challenge they faced with this type of dynamic imaging of synthetic cells was poor signal-to-noise ratio (SNR). Dr. Deshpande shared, “We struggled with SNR due to the low quantum efficiency of our previous camera and the low exposure times of 10 ms, which are necessary to prevent photo-bleaching during imaging. We want to image with the lowest exposure time possible. We also want to image at 100 or 1000 fps, so sensitivity is really important.”
Dr. Deshpande told us that “Our previous camera also had a split sensor design which leads to degradation of image quality and image artefacts.”
We previously used to image in a single plane but now we can take z-stacks through the liposome with high enough frame rates to visualize the internal dynamics