Description:
X-Ray Assisted Electron Microscopy Staining Procedure
Princeton Docket # 19-3530
Researchers in the Princeton Neuroscience Institute, Princeton University have developed an X-ray assisted staining procedure, which allows for tight control over the staining process and can serve as a very important and unique entry point for staining quality control while the protocol is conducted. This could not only be very useful for any basic research lab that uses electron microscopy for example to study the ultra-structure of brain tissue, but more importantly, it could finally enable an industrial-scale, fully automated and tightly controlled electron microscopy staining pipeline, which will allow for a real- time, continuous check on the quality and homogeneity of the staining without damaging or destroying the sample. This could also be very useful for potential electron microscopy-based high-throughput drug screening assays in the pharmaceutical industry
Typical electron microscopy staining protocols for biological samples consist of several steps in which a sample (e.g. a piece of brain tissue) is immersed in various heavy metal solutions. These heavy metals react with and bind to lipids, proteins or DNA and thereby stain these macromolecules and cellular structures. In thick samples however, it is difficult to establish homogeneous staining through-out the sample, mainly because of different diffusion kinetics in different regions of the sample. So far, the quality and homogeneity of the staining could only be assessed after the whole staining protocol has been conducted, including a final embedding step in an Epoxy resin. A typical staining protocol takes several days until the usual assessment by electron microscopy is possible. That assessment involves cutting the sample into ultrathin sections that can be viewed in an electron microscope, which is a destructive and very time-consuming process.
Applications
• Electron microscopy staining for biological samples
• High-throughput drug screening assays
• Industrial-scale, fully automated and tightly controlled electron microscopy staining pipeline
Advantages
• Non-destructive, real-time quality control
• Uses commercially available equipment
• Live monitoring of staining quality and heavy metal diffusion
Intellectual Property & Development Status
Patent protection is pending. Princeton is currently seeking commercial partners for the further development and commercialization of this opportunity.
The Inventors
Adrian Wanner is a CV Starr fellow at the Princeton Neuroscience Institute. Wanner studies how populations of neurons compute and process information using a functional connectomics approach that combines functional imaging of populations of neurons with subsequent electron microscopy-based large-scale neuronal circuit reconstruction. He holds a master degree in natural sciences from ETH Zurich and a PhD in neurobiology from the University of Basel. In 2016 he was awarded with the Chiquet-Ehrismann Originality Prize by the Friedrich Miescher Institute (Novartis), Basel.
David Tank - David Tank is the Henry L. Hillman Professor in Molecular Biology, Professor of Molecular Biology and the Princeton Neuroscience Institute, and Co-Director of Princeton Neuroscience Institute. The area of Tank Laboratory is Computational and Systems Neuroscience. One of the major focuses of Tank Laboratory to develop methodologies and instrumentation that can provide measurements of chemical and electrical dynamics of neurons in vivo.
Sebastian Seung - Sebastian Seung is Anthony B. Evnin Professor in the Neuroscience Institute and Computer Science Department at Princeton University. Seung has done influential research in both computer science and neuroscience. Over the past decade, he helped pioneer the new field of connectomics, applying deep learning and crowdsourcing to reconstruct neural circuits from electron microscopic images. His lab created EyeWire.org, a site that has recruited over 250,000 players from 150 countries to a game to map neural connections. His book Connectome: How the Brain's Wiring Makes Us Who We Are was chosen by the Wall Street Journal as Top Ten Nonfiction of 2012. Before joining the Princeton faculty in 2014, Seung studied at Harvard University, worked at Bell Laboratories, and taught at the Massachusetts Institute of Technology. He is External Member of the Max Planck Society, and winner of the 2008 Ho-Am Prize in Engineering.
Contact: Laurie J Tzodikov at tzodikov@princeton.edu, 609-258-7256