LANL scientists develop 3D tracking microscope

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By John Severance

Jim Werner got the idea after attending a conference in Aspen, Colo., back in 2002.
“The conference highlighted two-dimensional views of a single idea and I got the idea we have to start doing this in three-dimensions,” Werner said this week. “I wrote a proposal and got it funded.”
In 2008, Werner and his staff at the Los Alamos National Laboratory came up with the world’s first confocal microscope capable of following the 3D motion of nanometer-sized objects, which earned a 2008 R&D 100 award. That award recognized the top 100 industrial innovations worldwide.
But Werner and his staff were not done yet.
Earlier his month, they developed a 3D tracking microscope to follow three-dimensional movement of individual protein molecules inside live cells.
In an early demonstration, this instrument was used to follow three-dimensional dynamics of key proteins involved in the human allergic response and associated biological signals.
The microscope system simultaneously samples four spots surrounding the molecule under scrutiny and tracks both its spatial and temporal dynamics. To facilitate such tracking, these important signaling molecules are labeled with quantum dots, tiny glowing nanocrystals.
The system enjoys several advantages over other approaches to 3D molecular tracking:
• an increased tracking range that enables detection of biomolecular motion throughout the entire volume of many mammalian cells
• substantially lower damage to the cell in which the molecules reside
• the ability to perform time-resolved spectroscopy on the molecules being tracked.
Werner and his 3D microcope tracking team of Peter Goodwin, Guillaume Lessard, and Nathan Wells work at the Center for Integrated Nanotechnologies, which is jointly operated by LANL and Sandia National Lab.
 “There are a handful of groups pursuing camera-based approaches,” LANL communications director Jeffrey Berger said. “These are limited to 1 micron in the applicable tracking range (called Z-tracking range) and suffer from large backgrounds. The methods used by Jim’s team work over 10 microns in Z-tracking range and are much better at tolerating large background, which could also be labeled “high-background mediums.”
“Two other groups--at Stanford and at Princeton--have demonstrated 3D molecular tracking using confocal feedback methods. Our group is the first to demonstrate this in live cells.”
Werner outlined the progression of the research.
“First we came up with the geometry and microscopic design and the first publication of our design was in 2006,” Werner said. “The simulations said it looked like it worked and in 2007, we got our research published. In 2008, we came up with the world’s first confocal microscope capable of following the 3D motion of nanometer-sized objects. In 2009, we did stuff on cell surfaces and then last year, we demonstrated we can track individual molecules inside of cells.”
 Werner has worked at the lab for the past 13 years, having done his post-doctorate work in 1997 and he became a technical staff member in 2002.
Werner went to high school in Albuquerque and earned his bachelor’s degree in applied physics at the California Institute of Technology and received his masters and PHD at Cornell.
“I wanted to work at the lab because I was originally from New Mexico and the project I was being recruited for was really cool and interesting,” he said. “They were working on sequencing DNA by single molecule detection.
“It was really vastly different than anything I was doing at Cornell and the payoff was really big.”
Apparently, it has been really big because Werner and his team keep climbing the ladder when it comes to this new innovative research.