Intellectual Merit: This DMR MRI award funded the acquisition of an Atomic Force Microscope (AFM) combined with a fluorescence microscope to support a growing need to probe and correlate the optical and electronic properties of bio-materials with the nanometer scale structure readily obtainable using AFM. Students and postdocs have used the instrument to study clathrin mediated endocytosis in macrophage obtained from mice. Endocytosis is a fundamental bio-physical process critically important in immune system response. The figure at right shows A) a graduate student inspecting a sample using the acquired inverted frame fluorescence microscope before scanning a fixed macrophage cell with the Asylum MFP-3D AFM (AFM shown on the stage of the optical microscope); B) The obtained topographic image showing endocytotic pits in the macrophage (dark spots), C) an AFM image overlaid on the corresponding fluorescence image of GFP-labeled clathrin, D) surface representation colored with fluorescence to indicate the presence of clathrin, E) zoom in image of clathrin coated pit, and F) line cut indicating the size of the pit (≈100nm). The work is a part of an NSF funded center (BioSNTR) focused on using imaging methods and bio- informatics to understand regulatory networks which control gene expression and cellular metabolic processes. SE nAN MRI: Acquisition of a Combined AFM and Fluorescence Microscopy System Steve Smith, South Dakota School of Mines and Technology, DMR A B A B C D E F

Broader Impacts: A) High school student Shaunak works regularly in the lab at SDSMT and has been trained in the use of atomic force microscopy. As part of his research experience, he used nano-fabrication techniques to fabricate nano- structured gold nano-triangle arrays, using nanosphere lithography. B) After fabricating these arrays, he imaged them using an AFM, the image shows ≈ 50nm nano-islands in a hexagonal array with a period of SE nAN MRI: Acquisition of a Combined AFM and Fluorescence Microscopy System Steve Smith, South Dakota School of Mines and Technology, DMR A B [1] S. Shende, J. Fisher and S. Smith, “The Production of a Nanotriangle Lattice for Use in Metal-Enhanced Upconversion of Light,” Proceedings of the South Dakota Academy of Science, Vol (2014). ≈ 500nm, C) As part of an outreach project, Shaunak used 3D- printing to replicate the images into 3D models, magnified 20,000X so that school age children could visualize nano-sized objects and even touch them. The work led to a first author published abstract in the South Dakota Academy of Sciences. C

SE nAN MRI: Acquisition of a Combined AFM and Fluorescence Microscopy System Steve Smith, South Dakota School of Mines and Technology, DMR Broader impacts: The PI and grad students Amy Sin Ping Hor and Jon Fisher, organized and hosted a fluorescence workshop during SDSMT’s annual GEAR-UP program (Gaining Early Awareness and Readiness for Undergraduates Program). The program is designed to increase the number of American Indian students that achieve success in higher education through a rigorous, pre- college enrichment program that is acceleration based and comprehensive by design The photos show the grad students working with Native American students to acquire and process fluorescence images (right) and construct a paper microscope from household materials (left).