Another Landmark Development in Biological Microscopy at the ALS (from ALS News Report, volume 299, June 24. 2008) The biotech, pharmaceutical, and biofuels industries all grew out of our ability to understand and then predict how cells respond to changes in their environment. For example, the quest to make biofuel production more efficient begins with understanding how the cells that carry out fermentation deal with increasingly toxic concentrations of alcohol or butanol, taking advantage of this knowledge to guide the development of organisms with increased tolerance toward these molecules. Microscopy is a key technology in such work, in particular, high-resolution three-dimensional methods such as soft x-ray tomography. The establishment of the National Center for X-Ray Tomography (NCXT) and the construction of XM-2 firmly put the ALS on the map as a premier facility for imaging cells. Recently, the ALS became home to a completely new bio-imaging method: cryogenic, high-numerical-aperture light microscopy.
June 24, 2009
Developed by Mark Le Gros and Carolyn Larabell of the NCXT, this new microscope satisfies a long-standing need in cellular imaging. Now, for the first time, it is possible to image a whole, hydrated cell at high spatial resolution using both light and soft x rays. Correlation of the two data sets allows visualization of detailed cellular structure (x rays) together with the location of molecules tagged with a fluorescent label (light). The combination of these two pieces of knowledge is the "Holy Grail" of cell biology and answers the fundamental questions of who, what, where, and when. In other words, which molecules are interacting, and where and when do these interactions occur in the cell. This is a very exciting development in the world of cellular imaging, with the ALS being the only facility in the world with this capability. This new multimodal imaging resource has already begun to be used to address a wide range of highly topical questions, ranging from the design of new drugs to fight malaria and fungal infections to understanding where biodiesel is stored in algae. The possible applications of this new technique are virtually limitless, and it opens up a new chapter in biological research at the ALS.
The new cryo-light microscope used in this work is described in detail in the paper, "High aperture cryogenic light microscopy," by M.A. Le Gros, G. McDermott, M. Uchida, C.G. Knoechel, and C.A. Larabell, to be published in the July 2009 issue of the Journal of Microscopy.
July 30, 2008
The Completion of XM-2 Heralds a New Era in Biological Imaging at the ALS
(Adapted from ALS News Report , volume 289, July 30. 2008)
Noted American physicist Richard Feynman once said "make the microscope one hundred times more powerful, and many problems in biology would be made very much easier." Four years ago, Carolyn Larabell, Mark Le Gros, and the staff of the National Center for X-Ray Tomography (NCXT) at ALS Beamline 2.1 took Feynman's words to heart and began the construction of XM-2, a new transmission soft x-ray microscope. This was the first such imaging facility in the world to be designed specifically for biological imaging. Fast forward to the present and the original vision has been realized. XM-2 is now fully commissioned and producing an unprecedented number of high-resolution three-dimensional tomograms of cells in their native state. Each imaging experiment only takes three minutes or less. The large field of view on XM-2 equates to relatively large numbers of cells being imaged in each experiment. For bacteria and yeast, this can vary from a handful to upwards of 40 tomograms being produced per experiment. Consequently, XM-2 has been prodigiously productive in the short period since it was commissioned. With existing techniques, such as electron microscopy, obtaining data to reconstruct a tomogram of even a single yeast cell is considered a Herculean achievement. With XM-2 this has become a relatively trivial task.
All of this beautiful work is now showing up in the literature, the most recent of which was the cover article of the Journal of Structural Biology (Parkinson et al., J. Struc. Biol. 162, 380 [2008]). This paper describes the unique characteristics of soft x-ray microscopy as a biological imaging tool. For the first time, the organelles inside a cell can be imaged quantitatively. This is essential for understanding the mechanisms that take place in normal cells and how they may change as a result of disease.
The next phase is the incorporation of other imaging tools into XM-2, the most notable of which is a high-aperture cryogenic light microscope (for which Larabell and Le Gros were recently awarded patent rights). This resultant multimodal imaging capability has created enormous excitement in the field. For the first time, it will be possible to image and identify fluorescently labeled molecules inside a cell then overlay this information onto a full soft x-ray 3D reconstruction. This revolutionary instrument and the developed correlated imaging methods ensure the ALS will remain at the forefront of biological and biomedical science for the foreseeable future and be a vital new tool for addressing the missions of NIH and DOE. For more information on becoming an NCXT user, click here.
June 2, 2008
NCXT's paper "Quantitative 3-D imaging of eukaryotic cells using soft X-ray tomography "on the cover of Journal of Structure Biology.
September 15, 2007
We collected the first tomographic data set from XM2!
(click on the picture to see the movie)
The sample is glass capillary coated with gold nanoparticles (single particle = 100nm). Data collected by Kristian Knoechel, Weiwei Gu and Mark Le Gros, and reconstructed by Dula Parkinson.
September 3, 2007
First image from XM2!
(Click on the picture for a larger image)
Taken by Mark Le Gros and Christian Knoechel, processed by Dula Parkinson.
March 31, 2007
NIH Director Elias Zerhouni visited the NCXT.
Read more (NIH Record, issue June 15, 2007).
We recently took delivery of a twenty node, eighty core parallel computing cluster of Apple Xserves using Intel Xeon processors. Using Open MPI and ten gigabit Myricom interconnects, it can reconstruct tomograms and perform other computationally intense operations in minutes instead of the hours it can take on a single processor.
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October 11, 2006
The ribbon was officially cut on the morning of October 11 to dedicate the new National Center for X-ray Tomography (NCXT). Located at Berkeley Lab’s Advanced Light Source, this new center features a first-of-its-kind x-ray microscope that will enable scientists to perform “CAT scans” on biological cells, just one of many unprecedented capabilities for cell and molecular biology studies.
