When a Nobel Prize winner writes a letter of recommendation for your grant application, it goes without saying that the funding agency usually takes a careful look. Such was the case for a grant awarded to Colorado State University from the National Institutes of Health that will allow Dr. John Rash to acquire a new tomographic transmission electron microscope for investigators in the College of Veterinary Medicine and Biomedical Sciences and other colleges at CSU, as well as provide the foundation to establish a National Center for Freeze-Fracture Replica Immunogold Labeling.
“I like to think that we helped Peter Agre obtain the Nobel Prize in Chemistry in 2003 through our contributions to his groundbreaking research findings on aquaporin water channels, and he has written a strong supporting letter to the NIH to continue the work that he pioneered,” said Dr. Rash, a Professor in the Department of Biomedical Sciences. “That work today is changing our very understanding of how cells function by showing us at a molecular level some of the fundamental processes of life.”
The $854,000 NIH grant, coupled with support from Colorado State University, will enable the purchase of a new tomographic TEM as well as a new freeze-fracture device that will serve as the catalyst to create the proposed National FRIL Center. Dr. Rash’s current laboratory is one of a few worldwide able to conduct research using the combination of TEM, freeze fracture, and advanced labeling techniques that allow researchers to examine at a molecular level the intricate structures of cells, including gap junctions, aquaporins, and ion channels; as well as visualize, identify, and label those molecules. Additional funding being sought from the NIH for the proposed FRIL Center will provide program support to help fund salaries, ensuring the long-term stability of the laboratory.
FRIL-labeled neuronal gap junctions
In freeze fracture, cells are quickly frozen in propane or by contact with a liquid nitrogen-cooled copper block, immobilizing cell components instantly. The block of frozen cells is fractured and surface ice removed. The fractured samples are shadowed by vaporized atomic platinum and a thin layer of vaporized carbon. Most of the remaining organic material is digested away and the replica is placed on a small TEM grid where it can be viewed using a tomographic transmission electron microscope.
As a potential national center, word of the laboratory’s capabilities has spread globally and collaboration requests are coming in faster than the lab is equipped to handle. With new research projects and collaborations at the Rash Laboratory expanding globally, the new equipment and funding courtesy of the NIH could not come at a better time. The laboratory’s current TEM is 20 years old and its freeze-fracture device is 22. The new TEM will have two digital cameras for improved high-resolution digital imaging, and a terabyte of storage for the complex images that take scientists deep into the molecular structures of cells.
“The improvement in resolution is somewhat like having a Hubble telescope to examine stars, while others are using a Coke bottle,” said Dr. Rash. “But, in this case, we are examining the individual molecules in cells. There are endless areas to explore and every place we touch we find something new; something really exciting that will end up changing our most basic understandings of how we function at the cell and molecular level.”