You are cordially invited to the Center for Biomolecular Structure Lecture 
Series,



Lanthanide-Binding Tags for 3D X-Ray Imaging of Proteins in Cells at Nanoscale 
Resolution



Lisa Miller, NSLS II Brookhaven National Laboratory

April 21, 2021, 13:30 (EST - NY time zone)

Link:  
https://bnl.zoomgov.com/meeting/register/vJItde2vrzotEhRC0VBwCs8cLqfFNJV3M3c





Abstract

X-ray Fluorescence Microscopy (XFM) is a powerful method for imaging the 
trace-element concentration, distribution, and speciation in cells and tissues. 
Even though the technique has been around for more than 30 years, only recently 
have advances in X-ray sources, optics, and detectors enabled two- and 
three-dimensional X-ray imaging at the nanoscale with attogram detection 
sensitivity. However, one limitation of XFM for imaging biological systems is 
detecting the trace-element distribution in the context of subcellular 
organelles and individual proteins. For visible light microscopy, the most 
ubiquitous method for imaging individual proteins within the context of a 
living cell is the use of intrinsically fluorescent proteins, such as the green 
fluorescent protein (GFP) that is co-expressed as a fusion tag along with the 
protein of interest.  However, visualization of these tags is limited by the 
wavelengths of visible light except by applying specialized super resolution 
approaches.  Here, we are developing applications enabled by encoded 
lanthanide-binding tags (LBTs), which are GFP-like analogs of minimal size (ca. 
15-20 amino acids) for XFM. In this talk, applications of LBTs to both 
membrane-bound and cytosolic proteins will be demonstrated in 2D and 3D using a 
15 nm X-ray beam. This approach enables visualization of LBT-tagged proteins 
while simultaneously measuring the elemental distribution in cells at a spatial 
resolution necessary for visualizing cell membranes and eukaryotic subcellular 
organelles.


The CBMS Team

-----------------------------

Vivian Stojanoff, PhD
Education, Training, Outreach
User Program
p 1(631) 344 8375
e nsls.lifescien...@gmail.com<mailto:nsls.lifescien...@gmail.com>
w https://www.bnl.gov/ps/lifesciences/<https://www.bnl.gov/ps/lsbr/>

Address:
Center for Biomolecular Structure
National Synchrotron Light Source II
Building 745
Brookhaven National Laboratory
Upton NY 11973



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