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Report on the use of Optical Tweezers to study the Organisation of Genomic DNA
Dr Rosalie Driessen is a post-doctoral researcher in the Laboratory of Molecular Genetics, part of the Leiden Institute of Chemistry at Leiden University. She is studying the organisation and dynamics of bacterial chromatin having received her doctorate from the University for her thesis entitled ‘The architects of crenarchaeal chromatin’.
Dr Driessen describes her research and talks about the ‘hows and whys’ of choosing to use optical tweezers in this work. “The research goal of our group is to understand how the genomic DNA is organised, in particular in bacterial and archaeal cells. Proteins that bind to the DNA (so-called nucleoid-associated proteins, chromatin proteins or architectural proteins) play an important role in dynamically shaping and compact the genome. These proteins are also referred to as architectural proteins as they shape the DNA by way of bridging, bending or wrapping the DNA. We want to understand how these proteins exactly function by understanding their DNA-binding properties and how they change the structure of DNA upon binding. In addition to conventional biochemical techniques, we conduct single-molecule experiments using techniques such as atomic force microscopy, tethered particle motion microscopy, magnetic tweezers and optical tweezers. Using optical tweezers (the JPK NanoTracker™) allows us to evaluate the physical properties of single DNA molecules (by pulling at single DNA molecules) and how this is affected by these specific proteins. We would like to use this in combination with fluorescent microscopy so we can actually visualise the proteins bound to the DNA and correlate this with the force response on the DNA molecule.” Dr Driessen works in the laboratory of Assistant Professor Remus Dame. Their on-going work is reported online at
http://molgen.lic.leidenuniv.nl/research/chromatin.
The JPK NanoTracker™ is a very powerful system. For the first time, dual beam force-sensing optical tweezers have been seamlessly integrated on inverted optical microscopes combining advanced optical and confocal techniques including single molecule fluorescence in a small footprint, easy to use system. JPK’s unique tweezers technology (also known as a Photonic Force Microscope) enables quantification of molecular, cellular and micro-rheological processes. Applications include molecular motor mechanics, binding/elasticity of DNA and proteins, cell membrane dynamics and particle uptake.
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New Software Provides Uncompromising Performance, Speed, and Usability for Raman, AFM, and SNOM
The new WITec Suite software is now available for all WITec imaging systems. It was specifically developed to acquire and process large data volumes of large-area, high-resolution measurements and 3D imaging while providing speed, performance, and usability. Through the software architecture and graphical user interface an integrated and consolidated functionality is available incorporating the various techniques and measurement modes from Raman, to AFM, to SNOM, fluorescence and luminescence.
An intelligent computer resource management provides the capabilities for the generation and visualisation of even large data sets. The high-speed data acquisition allows for example the measurement and recording of over 1300 Raman spectra in only one second. Furthermore data sets including several million image pixels, each containing the information of e.g. a complete Raman spectrum or an AFM pulsed force mode-curve, can be generated, processed, and imaged smoothly with WITec Suite.
Another focus of WITec Suite is the improved and simplified usability. The software design provides a clear and intuitive menu guidance and an individually adjustable user interface to be suitable for all experience levels and user requirements. The smart access options for all principle functions, including for example the circle mouse menu, accelerate the workflow and smooth the first steps into the software and an accessible learning curve.
“WITec imaging systems are well-known for their exceptional imaging qualities. Unprecedented performance and speed facilitate the acquisition of large data volumes and the generation of 3D images and large-area scans,” explained Dr Olaf Hollricher, R&D Director at WITec “The capabilities of WITec Suite match perfectly with the requirements for high-speed data acquisition and processing of large data volumes and provide an accomplished combination of comprehensive data analysis and ease-of-use.”
WITec Suite includes Control FOUR, a powerful software tool for measurement control and data acquisition, and Project FOUR, a user friendly data evaluation and processing software. The license terms facilitate the installation of Project FOUR on an unlimited number of computers permitting the user to process data and generate images wherever required.
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