AUG 13, 2019 9:00 AM PDT

Nanometer-accuracy distance measurements between fluorophores at the single-molecule level

Sponsored by: Andor
C.E. Credits: P.A.C.E. CE Florida CE
  • Research Specialist - UCSF/HHMI
      Nico Stuurman is a Research Specialist at the University of California, San Francisco, in the lab of Ron Vale. Nico obtained his Ph.D. at the University of Amsterdam in the Netherlands, studying organization of the cell nucleus. He then studied Drosophila nuclear lamins at SUNY, Stony Brook and at the Biozentrum, Basel Switzerland (with Ueli Aebi). He has a long standing interest in the use of computers and automation in the lab, and wrote an application to interface a multi-well plate reader with an Apple II computer in the 1980s. He also is interested in light microscopy, and combined these interests to advance many projects including the Open Source software, Micro-Manager . He taught at several courses (MBL Physiology, AQLM, Bangalore Microscopy course), and some of his didactic microscopy lectures are available through ibiology (
    • Imaging Application Specialist, Andor
        Dr. Barszczewski is a Belfast-based research applications scientist and training specialist at Andor Technology. He has 15 years' expertise in a wide range of imaging applications for CCD, EMCCD, and CMOS cameras as well as diverse microscopy and image-data visualization systems. He obtained his Ph.D. in neuroscience from the Max-Planck Institute for Biophysical Chemistry and the University of Goettingen, Germany in the lab of Reinhard Jahn.

        He has held various technical roles at Andor, now an Oxford Instruments company, advising customers in areas of astronomy and low-light biomedical imaging as well as physical imaging, including X-ray and quantum imaging, and advanced image-data visualization and analysis. He has been involved in the creation of application notes and technical white papers on established and emerging imaging applications, trends, and technologies. In addition, he continues to be actively involved in supporting pan-European and global research projects within the European Union's Framework Programmes. Dr. Barszczewski's specialties include biomedical, low-light and single-molecule imaging, molecular neuroscience, and scientific writing.

      DATE:   August 13, 2019
      TIME:    9:00am PT, 12:00pm ET, 5:00pm BST
      Molecular complexes are major constituents of cells, hence unraveling their mechanisms is key to fuller comprehension of cell biology. Electron Microscopy and X-ray crystallography provide high resolution static images, but deeper understanding requires dynamic measurements.
      During the past few decades, we have gained the ability to place small molecules (such as fluorescent probes) on specific sites of molecular machines. However, while single-molecule Förster resonance energy transfer (FRET) can determine relative distance changes between ~2-8 nm, direct distance measurements are difficult to achieve in part because FRET depends on dye orientation and requires careful calibration. Other current super-resolution measurements become error-prone below 25 nm. We developed methods for reliable image registration, and used available information about localization accuracy to measure average distances between fluorophores at sub-nm precision. These procedures can now be used to measure distance between multiple sites of a (protein) complex in different conditions.
      This webinar will explain the procedures for sub-nm image registration, explain how distance measurements can be improved by using localization precision information, and show how these procedures can be extended to dynamic distance measurements of biological nano-machines.
      Learning Objectives:
      • Recognition of the potential of "standard" microscopy equipment to deliver nm precision measurements between single fluorophores
      • Awareness of the impact of local image distortions on registration of multi-channel images of single molecules
      • Understanding the value of the precision of localization for individual molecules and its impact on distance measurement
      Webinars will be available for unlimited on-demand viewing after live event.

      LabRoots is approved as a provider of continuing education programs in the clinical laboratory sciences by the ASCLS P.A.C.E. ® Program. By attending this webinar, you can earn 1 Continuing Education credit once you have viewed the webinar in its entirety.

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