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Laser Analytics Group


'Democratising high-quality live-cell super-resolution microscopy enabled by open-source analytics in ImageJ'

Ricardo Henriques (UCL)

In this talk I will present high-performance open-source approaches we have recently developed to enable and enhance optical super-resolution microscopy in most modern microscopes, these are NanoJ-SRRF, NanoJ-SQUIRREL and NanoJ-Fluidics. SRRF (reads as surf) is a new super-resolution method capable of enabling live-cell nanoscopy with illumination intensities orders of magnitude lower than methods such as SMLM or STED. The capacity of SRRF for low-photoxicity, allows unprecedented imaging for long acquisition times at resolution equivalent or better than SIM.  For the second part of the talk, I will introduce SQUIRREL, an analytical approach that provides quantitative assessment of super-resolution image quality, capable of guiding researchers in optimising imaging parameters. By comparing diffraction-limited images and super-resolution equivalents of the same acquisition volume, this approach generates a quality score and quantitative map of super-resolution defects. To illustrate its broad applicability to super-resolution approaches, we demonstrate how we have used SQUIRREL to optimise several image acquisition and analysis pipelines. Finally, I will showcase a novel fluidics approach to automate complex sequences of treatment, labelling and imaging of live and fixed cells at the microscope. The NanoJ-Fluidics system is based on low-cost LEGO hardware controlled by ImageJ-based software and can be directly adapted to any microscope, providing easy-to-implement high-content, multimodal imaging with high reproducibility. We demonstrate its capacity to carry out complex sequences of experiments such as super-resolved live-to-fixed imaging to study actin dynamics; highly-multiplexed STORM and DNA-PAINT acquisitions of multiple targets; and event-driven fixation microscopy to study the role of adhesion contacts in mitosis.

Tuesday, 26 February, 2019 - 14:00 to 15:00
Contact name: 
Karen Scrivener
Contact email: 
Contact phone: 
+44 (0)1223 761208
Event location: 
Johnson Matthey Lecture Theatre (LT2), Department of Chemical Engineering and Biotechnology, Philippa Fawcett Drive, Cambridge CB3 0AS