The National Health Research Institutes (NHRI) of Taiwan and Asylum Research are pleased to sponsor the first Taiwan Bioworkshop. The Taiwan Bioworkshop is organized for scientists and students to share and exchange AFM research that is being done in the field of life science and biology. The Bioworkshop will combine lectures from leading researchers and industry experts, as well as instrument demonstrations using the Asylum Research MFP-3D™ AFM. This free event is open to all researchers in the field of AFM.
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The Taiwan Bioworkshop will be held 30-31, July 2009. National Health Research Institutes at Zhunan Campus David Beck, Asylum Research The Taiwan Bioworkshop will cover a variety of topics which include: • Principles of AFM Program Download meeting program
Friday, 31 July Instrument Demonstration Applications Scientists will instruct small groups of researchers (six to eight students) on a variety of AFM topics. Instruction will be done on the MFP-3D™ AFM. Attendees may sign up for a maximum of two topics when registering. 1:00-2:30 2:45-4:15 Principles of Atomic Force Microscopy This presentation will focus on the principles of AFM including instrumentation hardware, software, different scanning modes, and AFM basics for biologists. Force Measurements AFM can be used to probe nanomechanical and other surface properties such as adhesion and elasticity on biological samples such as DNA, cells, etc. AFM measures forces on the probe tip as it approaches and retracts from the surface, elucidating local mechanical and chemical properties. This presentation will cover the basics of force measurements, how it works, how to interpret a force curve, and instrumentation needed for low noise measurements. Biological application examples will be illustrated. Atomic Force Microscopy in Biomedical Studies Atomic Force Microscope (AFM), one of the most powerful research tools in nanotechnology, has become increasing important in biological and biomedical research recently. Although enjoying similar degree of resolutions, the AFM have many advantages over the electron microscopy. The sample preparations for the AFM imaging are relatively simple; no harsh physical or chemical treatments are required. Thus, the disruption of the samples during the preparations is minimised. Among all, the most important feature of the AFM is the fact that it permits the observation of samples in buffer solutions, so that biological samples can be studied at nanometre scales in their native and functional states under their physiological conditions, allowing not only their structure, but also their dynamics to be analysed. This presentation will demonstrate some of the applications of the atomic force microscopy on the biomedical studies we carried out, including phosphorylation measurements on biological membranes, analysis of photocatalytic treatments on bacterial cells, detailed imaging of bactericidal effects of antimicrobial peptides, morphological studies of amyloid-beta aggregation which is normally linked to Alzheimer disease, as well as the imaging of hepatitis C virus core auto-assembly. Biological Applications and Sample Preparation for AFM Integration of AFM with Optical Techniques AFM is an indispensible tool for high resolution imaging and measurement of topography and mechanical properties of cells. Coupled with the optical microscopy’s ability to label and identify cellular components, these two complementary techniques provide a complete picture on cellular structure and mechanics, not readily available with each technique individually. This tutorial will discuss: 1) hardware requirements for integration without interference during realtime acquisition of both AFM and optical images, 2) software features necessary to direct the AFM within the optical window to capture either an image, individual force curves or force map, and 3) offline software capabilities for AFM and optical data integration, image analysis and image overlay, including 3D rendering. Relevant examples will be presented. Platforms for Cell Mechanics Evaluation Tung-I Lin1, Ying-Ting Chen2, Meng-Ru Shen3, Jen-Fin Lin1, Hsien-Chang Chang1,2 Mechanical force plays a critical role in the interactions of cell with their surrounding extracellular matrix (ECM). These processes are critical for control cell growth, migration, differentiation, and apoptosis during organogenesis and wound repair [1]. Here we utilized AFM to evaluate the mechanical behavior of cell-cell junction of wild-type/EMT(STIM1, KCC3 overexpression) SiHa cervical cancer cell line through mathematical model created by depth-sensing indentation of composite materials. We also studied the role of the calcium sensor molecule within endoplasmic reticulum: STIM1 on cervical cancer cell migration by the real-time, high-resolution measurements of calcium spikes, molecular trafficking and traction force applied by cells at single adhesion sites. The novel approach combines a variety of techniques, including AFM, development of mPADs [2], confocal/multiphoton fluorescence imaging of Ca2+ flux and focal adhesion complexes in living cells. The AFM preliminary results between wild/EMT type SiHa cells indicate the Young’s modules change of cell-cell junction was due to the cytoskeletons distribution within the cellular edge, these were correlated with cell surface topography by AFM. We also demonstrate that STIM1, which is involved in store-operated calcium entry, is essential for cervical cancer cell migration. The molecular mechanism by which Ca2+ influx regulates cell migration at least partly involves the modulation of focal adhesion turnover, hereby affecting cell traction force. Acknowledgement: Reference
Gramicidin is a linear antibiotic ion-transport-channel peptide that forms two different dimer conformations depending on the environment: a single stranded helical dimer, or a double stranded anti-parallel helical dimer. These structures provide the mechanism and pathway for transporting small (<0.5nm) monovalent cations through bacterial membranes, thus changing the ion gradient between the inner and outer environment and accounting for gramicidin’s antibacterial activity. The gramicidin antibiotic peptide is known to form aggregates in one of two states in lipid bilayer (LB) experiments. When the films are deposited on surfaces such as mica and graphite, these molecules may exhibit either a flat lying or standing up orientation. However, in biological systems, gramicidin molecules often encounter membranes and other surfaces which are not well-modeled by a cellular lipid structure. In these cases, gramicidin aggregation and orientation behavior is not well understood. In this presentation, we show how gramicidin molecular film formation and aggregation may be controlled by the density of the molecules (inter molecular forces) analogous to pressure control in LB films. We further show how the substrate chemistry and temperature can very strongly influence the film formation and molecular orientation on the surface. Finally, we show that by careful control of these parameters, nanoscale features and structures can be created. Cell Mechanics Using Atomic Force Microscopy: Adhesion, Indentation, and Compression Future Directions of AFM in Biological Research This presentation will discuss the latest research being done in AFM in biology. A roundtable discussion will follow. The Bioworkshop is free to all attendees. Lunch will be provided both days. All attendees must register for the conference and may select two topics in the instrument demonstration sessions. The following hotel is recommended and is located near the NHRI campus: Travel information to the NHRI campus can be found here. -Maps For additional information, please contact:
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