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Direct Imaging of Aligned Neurofilament Networks Assembled in Microchannels Neurofilaments (NFs) are major constituents of axons and dendrites of vertebrate.

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Presentation on theme: "Direct Imaging of Aligned Neurofilament Networks Assembled in Microchannels Neurofilaments (NFs) are major constituents of axons and dendrites of vertebrate."— Presentation transcript:

1 Direct Imaging of Aligned Neurofilament Networks Assembled in Microchannels Neurofilaments (NFs) are major constituents of axons and dendrites of vertebrate nerve cells (Figure, Left). In vivo, these filamentous proteins assemble to form networks that act as scaffolds for microtubules and organelles, and provide structural and mechanical stability to neuronal processes (axons and dendrites). In-vitro studies show that NF-networks form a novel liquid crystalline hydrogel (Biophysical Journal 2008, 95, 823- 835) (LEFT) AFM phase image of isolated neurofilaments deposited on polyethyleneimine (PEI) coated silicon wafer. Inset show height image with the same topographic features. Image size is 1  m 2. Inset size is 500nm 2. (Adapted from Langmuir 2008, 24 (16) 8397- 8401) (RIGHT) Cover Image for August 19, 2008 issue of Langmuir: From macro to nano scale imaging of aligned neurofilament (NF) networks in microchannels. The microchannel in situ dialysis chamber device is shown on the mm scale. Cross polarized optical microscopy images of NF hydrogel formed in 50x50  m 2 channels is shown on the 250 micron scale. Images were taken at 45 degree to analyzer showing bright spots comprised of oriented NF-networks. Nanoscale imaging using AFM phase contrast on aligned NF-networks inside a 50  m wide microchannel. Right image size is 1.5  m 2. Left image close up is 300 nm 2. Cyrus R. Safinya, UC Santa Barbara, DMR 0503347 & 0803103 We have developed a micro-fluidic based method to produce aligned neurofilament networks for direct imaging and structural studies in a microfluidic device. The alignment was achieved by assembling neurofilaments from protein subunits confined within microchannels. The resulting network structure was probed by polarized optical microscopy and atomic force microscopy, which confirmed an unprecedented degree of long-range NF protein alignment within microchannels (Figure, Right). This technique is expected to enable structural studies of hierarchical assemblies of a range of biopolymer assemblies (proteins, nucleic acids) paving the way for the understanding of the nature of their intermolecular interactions under biologically relevant conditions. (Langmuir 2008, 24 (16), 8397-8401)

2 Education and Outreach Research Training: A Biomolecular Materials Emphasis Cyrus R. Safinya, UC Santa Barbara, DMR 0503347 & 0803103 Education: Multidisciplinary teams comprised of undergraduate and graduate students, and postdocs, with backgrounds in materials science, physics, chemistry, and biology, are educated in methods to discover nature’s rules for assembling the molecular building blocks in distinct shapes and sizes for particular functions. The learned concepts enable development of advanced nanoscale materials for broad potential applications in electronic, chemical, and pharmaceutical industries. Outreach: Katelyn Cahill Thompson (top photo, right) is studying to become a biomedical engineer at UC Davis. To date, she has successfully completed her sophomore year of college at UCD. She is currently a RISE (research internship in science and engineering) intern at UCSB (summer 2008), and together with her mentor graduate student Joanna Deek (top photo, left) is working on a project involving the self-assembly of neurofilament subunits under various biologically relevant solution conditions. Daniel Orosco (middle photo, right) is an undergraduate Pre-Med senior in the Molecular, Cellular and Developmental Biology department at UC Santa Barbara. He is supported by the California Alliance for Minority Participation (CAMP), which aims to promote diversity of undergraduates in research. Daniel is working in the PI’s lab with his mentor graduate student Nate Bouxsein (middle photo, left) on improving the transfection efficiency of cationic lipid vectors for gene delivery and silencing. He is testing a library of cationic lipids complexed with plasmid DNA, small interfering RNAs (siRNA) or short synthetic DNAs. The goal is to determine how the length, structure and design of the nucleic acids effect the stability of the various lipid complexes. Exchange undergraduate student Sophia Rudorf (bottom photographs, left), from the University of Potsdam in Germany initiated experimental work in the PI’s laboratory during the Winter and Spring 2008 quarters. Sophia studied the interactions between DNA molecules adsorbed on supported lipid bilayers with tunable membrane charge density. She was mentored by Postdoctoral Fellow Cecilia Leal (bottom photographs, right) and was trained on how to use x-ray scattering (bottom right photo) and optical fluorescence microscopy (bottom left photo).


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