JPK's ForceRobot system
produces high volume single molecule force
spectroscopy results at the University of Nebraska

JPK Instruments, a world-leading manufacturer of nanoanalytic instrumentation for research in life sciences and soft matter, reports on the work from the Pharmaceutical Sciences Department of the Medical School at the University of Nebraska. The Department has selected the JPK ForceRobot® 300 system to extend their studies applying atomic force microscopy, AFM, in the measurement of single molecule force spectroscopy.

Dr. Alexey Krasnoslobodtsev working with the JPK ForceRobot® 300 system.

Professor Yuri Lyubchenko heads a research group at UNMC in genomics. Their goal is to unravel the role of the DNA dynamics at different levels as key mechanisms for various DNA functions including gene regulation, DNA recombination and the mismatch repair with the major focus on DNA recombination. The misfolding and aggregation of proteins are widespread phenomena leading to the development of numerous neurodegenerative disorders for which there is currently no cure.

Lyubchenko's group has published widely using AFM as both a direct and complimentary tool to advance their understanding of biological processes, some of which are relevant to human health. Single molecule force spectroscopy is used to study interactions between peptides and proteins involved in the development of these neurodegenerative diseases such as Alzheimer's and Parkinson's diseases. AFM helps to bridge the gap of various techniques which are capable of looking at the complex processes of protein misfolding and aggregation. However, it has the drawback of being very time and labor consuming to make measurements. This has been solved by the introduction of JPK's ForceRobot® 300 system into the group.

One of the key researchers in the group is Dr. Alexey Krasnoslobodtsev. He did his first degree in Chemistry at Novosibirsk State University before completing his PhD at the New Mexico State University. Speaking about his research, Krasnoslobodtsev says "AFM is my instrument of choice in my work towards a better understanding of these protein misfolding and aggregation phenomena. By measuring strength of interactions between protein molecules, it is possible to detect pathological misfolded conformations of proteins which are capable of triggering aggregation. Such conformations are characterized by elevated propensity to interact with one another. It is hoped that better understanding the mechanisms underlying self-assembly of proteins and peptides into nano-aggregates of various sizes and morphologies would eventually facilitate the development of efficient therapeutic and diagnostic tools for diseases associated with protein misfolding."

He continues: "The first time I saw a presentation about ForceRobot was at the Biophysical Society meeting in 2007. I remember sharing my excitement with my colleagues in the lab after I got back. The ForceRobot seemed to have the ability to take over the tedious routine manipulations like data acquisition and analysis leaving more time for conceiving ideas and thinking about science. Although, much is still in the hands of the researcher, for example sample preparation and data interpretation, automated data collection makes it much easier to carry out an experiment. For example, when I come to work in the morning and find ForceRobot (which was left running overnight) is about to finish collecting thousands of force curves, I feel really excited because I can look at the data right away."

JPK develops, engineers and manufactures instrumentation in Germany to the world-recognized standards of German precision engineering, quality and functionality.

For further information of the ForceRobot® 300 and other products in the JPK family of nanoscale characterization instrumentation, view website: www.jpk.com or Facebook: www.jpk.com/facebook  Refer to page 228 

Linkam tensile stage chosen to control stress in
growth of human collagen by the University of Liverpool

Market leaders in temperature controlled microscopy, Linkam Scientific Instruments, have been chosen by the Physics Department of the University of Liverpool to study the effects of controlling stresses applied during the growth of collagen secreted from human fibroblast cells.

Picture shows Dr Smith's experimental set up: a PDMS well mounted in the LinkamTST350 with Fibroblast Cells in media.

The research goal of Senior Research Assistant, Dr Caroline Smith, and her colleagues at the University of Liverpool is to advance the treatment of tendon injuries by developing a method of growing oriented human collagen.

To achieve this and to gain a fundamental understanding of the process, Dr. Smith is using the optical technique of reflection anisotropy spectroscopy to monitor the extent to which collagen, grown by mouse fibroblast cells on elastomeric substrates, is oriented when subjected to a regular uniaxial stress mounted in a Linkam TST350 tensile stage.

For the experiment, small wells capable of holding liquid were fabricated from polydimethylsiloxane (PDMS) to fit in the Tensile Stress Tester. The effect of stretching the PDMS as a function of applied stress was investigated initially to determine the effect of stress on the substrate. (J. Phys. D: Appl. Phys. 43, 245301 (2010)). L929 murine fibroblasts cells were cultured at 37˚C in a 5 % CO2 atmosphere in Dulbeccos Modified Eagles Medium (DMEM) (Sigma) supplemented with 5 % Foetal Bovine Serum (Invitrogen) and 1 % Penicillin/Streptomycin antibiotics (Sigma). The culture medium was replenished daily during the cycling period. The cells were then subject to cyclic stress for 7 days. After 7 days, the wells were removed from the incubator and washed with Dulbecco's Phosphate Buffered Saline (DPBS, Sigma). The cell layer was dispersed in 0.05 % Trypsin/EDTA (Sigma) incubated for 3 min and subjected to gentle agitation to remove the cells without damaging the collagen layer. After removal of the cells the collagen secreted from the cells could be investigated by optical microscopy and reflection anisotropy spectroscopy.

The ability to choose a wide range of cyclic speeds to relate back to the growth of collagen is very useful. Initially, Dr Smith thought about making her own TST but having a small unit that could control the cyclic stress and also measure stress/strain curves at the same would be very difficult to manufacture in-house. Hence, the Linkam unit was purchased.

The TST350 stage is built with two precision ground stainless steel lead screws to maintain perfect uniform vertical and horizontal alignment. The sample jaws move in opposite directions to maintain sample in both reflected and transmitted microscope fields of view. Temperature control and accuracy is excellent with a range from -196°C to 350°C with 0.01°C control and up to 30°C/min rates, with virtually no temperature feedback to the measurement of force. Samples can be quickly loaded into the jaws and a test run can be performed in seconds. The data feedback from the force transducer, designed and built in-house, can be used to display an online plot of the force/distance when the jaws are moving at constant speed or when speed is varied to maintain a constant force. The speed of the jaws, force applied and distance moved can all be varied relative to the cell temperature. The sample chamber is sealed and can be controlled with various inert gases via the precision click fit valves built onto sides of the stage.  Refer to page 230

For further information, view website: www.linkam.co.uk today and learn about the broad range of applications in the field of temperature controlled microscopy.

Thermo Fisher Scientific's new method for low
level detection of volatile Nitrosamines in tobacco

Thermo Fisher Scientific's offer a new comprehensive method, using triple quadrupole GC-MS/MS, to achieve lower levels of detection of volatile nitrosamines (VNA) in tobacco. The new method allows environmental laboratories, tobacco companies and government agencies to efficiently separate VNAs while lowering detection limits, increasing specificity and enabling the analysis of many other contaminants in tobacco, including pesticides. The new method is detailed in an application note entitled "Lower Detection Limits of Volatile Nitrosamines in Tobacco by Triple Quadrupole GC-MS/MS," available to download a: www.thermoscientific.com/vna

Volatile nitrosamines are a class of compounds that can form in tobacco smoke, as well as during the curing and processing of tobacco. These compounds have been proven to have adverse effects on human health; and two of the VNAs found in tobacco, N-nitrosodiethylamine (NDEA) and N-nitrosodimethylamine (NDMA), are classed as known human carcinogens by regulatory authorities. As a result, these compounds must be thoroughly monitored in order to safeguard human health and comply with increasingly stringent regulations.

The new Thermo Fisher method is a powerful alternative to other traditionally used techniques. By coupling a gas chromatograph to a triple quadrupole mass spectrometer, the new method makes detection limits of 1ng/mL achievable, enabling users to satisfy the increasingly lower detection limits required by governments and regulatory bodies. The method also increases contaminant specificity within a classification and allows many other organic contaminants and chemicals in tobacco to be analyzed simultaneously, including pesticides.

The new method includes GC-MS/MS analysis on the Thermo Scientific TSQ Quantum XLS triple quadrupole GC-MS/MS system using timed selected reaction monitoring (t-SRM). This unique feature enables easy method set-up and allows users to run samples while the instrument automatically determines the optimal time for SRM parameters. For further information, e-mail: analyze@thermofisher.com or view website: www.thermoscientific.com/gc

Alto Marketing reappointed as Media Relations Partner
for European Laboratory Automation (ELA) for second year

Following the success of Select Biosciences' first European Laboratory Automation (ELA) in 2011, Alto Marketing, the specialist Life Science and Healthcare communications consultancy, is pleased to announce that it will be conducting the PR programme and running the on-site press room for the second consecutive year. Taking place at the Congress Centre Hamburg (CCH), Germany, 30-31 May 2012, the event offers a comprehensive schedule of informative and topical presentations, workshops and training sessions focused on automated techniques, equipment and applications.

The first ELA attracted 1523 attendees and 151 exhibitors and media partners. Ken Browne, Event Organiser and Director of Select Biosciences commented, "We were delighted with the success of the first ELA conference and exhibition and received some fantastic feedback. We are aiming to fill the need for a pan-European Automation event, and establish ELA as the forum for combining scientific content with up-to-date technology; having Alto Marketing as our PR partner is part of our strategy for achieving this."

Registration for ELA 2012 is open now and delegates have the option to attend 8 different conference streams, each of which includes internationally renowned keynote speakers, research presentations, training sessions and much more.

David Robinson, Managing Director of Alto Marketing remarked, "We're really pleased to be partnering with Select Biosciences on ELA again in 2012, as it ties in so well with a number of our other clients. We will be working with the ELA team closely over the coming months, providing event support PR to build on the success of 2011 and help maximise delegate and exhibitor attendance. Managing the media room on site means that we can also service the needs of the attending press. It's a fantastic event for the media, and we look forward to welcoming new editors and journalists in 2012, in addition to those that attended this year."

Established in 2002, Alto Marketing is a committed and forward-thinking specialist healthcare and life science communications consultancy.

With extensive experience of worldwide marketing in sectors including life science research, (bio)pharmaceuticals, diagnostics and medical devices, Alto Marketing develops and implements communications strategies for diverse clients ranging from start-up technology companies to multi-billion dollar global corporations.

Alto's marketing philosophy is based on four pillars:
1) Alto Advise: The consultancy and strategy service
2) Alto Message: The marketing content development service including technical science and healthcare writing, scientific business writing and sales and promotional content development
3) Alto Announce: The service to publicise marketing messages through public relations, advertising and direct marketing
4) Alto Engage: The personalised and interactive offering covering social media, email marketing and webinars.

The company is dedicated to providing full accountability and outstanding project management and direct business results.

For further information on Alto Marketing's unique communications expertise in Life Science and Healthcare, view website: www.alto-marketing.com