Bitplane Imaris 4D Software helps the Centre
for Microvascular Research understand regulation
of transendothelial migration of neutrophils in vivo

The migration of neutrophils from the vascular lumen into inflamed tissues is a crucial component of the normal immune response and a significant factor in the development of inflammatory pathologies, where inappropriate inflammation contributes to tissue damage and disease progression. A decisive step in this process is the passage of the blood neutrophils through the endothelial cells lining the lumen - transendothelial migration (TEM).

The complex structure of the vessel wall and the contribution of environmental factors, such as blood flow and local chemokine production, have limited the value of in vitro studies into the mechanisms governing this process. Now, advanced Imaris 4D modelling software from Bitplane is enabling in vivo analysis of the dynamics of leukocyte migration with a high degree of spatial and temporal resolution, leading to a new understanding of TEM regulation highlighting the role of junctional adhesion molecule C (JAM-C).

A luminal view of a leukocyte migrating through the junction between three endothelial cells to the sub-endothelial space. Other leukocytes can be seen crawling on the luminal surface of the venule.

The work is reported in a paper by Dr Abigail Woodfin and her colleagues at the Centre for Microvascular Research, which investigates the molecular and cellular events within the microvasculature focusing on the regulation of vascular integrity, morphology and function. The Centre has a strong and internationally acknowledged expertise in the application of specialised imaging methods, including confocal intravital microscopy which allows in vivo observation of events within the microcirculation, in 3D, in real time.

"After investigating several 3D modelling platforms we chose Bitplane Imaris. As well as a comprehensive feature set and user-friendly interface, it proved to be the most stable when analysing very large files, which was essential to our work," says Dr Woodfin. "Because of this, Imaris enabled us to track the movement of leukocytes relative to the endothelial cells lining the vessel wall over many sequential time points.

"During long periods of analysis there is always a degree of movement with living tissue and we used the Imaris drift correction function to correct for this. The crucial thing was that Imaris allowed us to convert our data into a virtual 3D object that could be fully manipulated in terms of rotation position, zoom, and the intensity at which each channel is displayed. This is not possible using 2D projections of Z-stacks, which is how confocal microscopy images are traditionally presented."

A low magnification view showing a post capillary venule and capillaries in the mouse cremaster. Leukocytes can be seen crawling on the luminal surface of the vessel, and migrating through the extravascular tissues.

A low magnification view showing a post capillary venule and capillaries in the mouse cremaster. PECAM-1 is enriched at the junctions between endothelial cells and expressed at lower levels on the body of the endothelium.

"Fast, precise and easy-to-use, Imaris is a uniquely powerful and versatile solution for the visualization, analysis and interpretation of 3-D and 4-D images," says Marcin Barszczewski at Bitplane. For additional flexibility, Imaris also offers a range of eleven specialist modules, including MeasurementPro, Track, Coloc and FilamentTracer."

For further information, view website: www.bitplane.com

Thermo Fisher Scientific makes it easy to match the right plate to your assay

The new interactive online Thermo Scientific immunoassay plate guide. provides a detailed resource for users looking to match the right plate to each assay. With information on passive binding surfaces for biomacromolecules, covalent coupling surfaces for smaller biomolecules and affinity capture surfaces for affinity-tagged biomolecules, you can be confident that you are using the best plate for each application.

Immunoassay techniques are extremely sensitive, making it pivotal that you select the appropriate surface with optimized conditions. The portfolio of Thermo Scientific Nunc plates provides a broad range of surfaces and formats to optimize your assay.

For further information, and to view the plate selector guide, view website: www.thermoscientific.com/oemdiagnostics  Refer to next page

New device mitigates spectral performance
issues related to temperature variation

To maintain thermal stability of spectrometers operating in variable environments, Ocean Optics has released the USB-TC small-footprint heating device. Compatible with the company's USB2000+ and USB4000 miniature spectrometers, the unit attaches directly to the spectrometer and maintains wavelength thermal stability to within +/-0.025 pixels/ ºC. This temperature control ensures accuracy in industrial and other settings where temperature variations can affect spectral performance.

The USB-TC is available with pre-selected setpoint temperatures between 25-55ºC and comes with a 12 VDC, 3A power supply. With constant ambient temperature, the USB-TC stabilizes to within 0.1ºC of the final temperature within 30 minutes of power-up. The system will operate in temperatures ranging from 5-40ºC below the setpoint temperature.

The USB2000+ and USB4000 are among the most popular miniature spectrometers in the world. Their small-footprint, high-speed CCD-array design is ideal for a wide range of applications. Modular optical bench accessories (entrance apertures, order sorting filters and more) make it possible to configure the USB2000+ and USB4000 for hundreds of different applications.

The USB-TC is available with new spectrometer purchases or as a retrofit option for existing owners of USB2000+ and USB4000 spectrometers.

For further information, view website: www.OceanOptics.eu  Refer to next page

Olympus BX3 systems with true colour LED illumination and Light
Intensity Manager for comfortable and efficient clinical microscopy

Olympus provides its versatile, powerful BX3 clinical microscope systems for analysis and disease diagnosis in pathology and cytology. The systems take advantage of Olympus's new true colour LED illumination technology and built in Light Intensity Manager (LIM) to create images with accurately rendered colours. Histological and cytological stains appear exactly the same under the true colour LED as they do under daylight filtered halogen, facilitating a seamless transition for labs adopting the new, more efficient LED technology. True colour LEDs offer longer lifetimes, constant colour temperature at all voltages and reduced power consumption when compared to traditional halogen bulbs. In addition, the LIM improves user workflow and maximises consistency by automatically modulating light intensity when working with different magnifications. Such advantages make the BX3 systems with true colour LED illumination the logical choice for comfortable and efficient clinical microscopy.

Insightful analysis in pathology and cytology hinges upon the years of experience provided by histological experts, developed using traditional halogen bulbs in combination with daylight filters. LEDs provide clear energy saving benefits but until now have not generated the level of colour reproduction obtained using halogen approaches. The Olympus true colour LED approach resolves this dilemma by utilising the most advanced mixed-matrix brightfield LED technology currently available to provide a colour rendering index very similar to that of halogen illumination. Stains such as Haematoxylin & Eosin and Papanicolaou look the same when using the Olympus LED as when using halogen light sources, while similar colours can be easily differentiated. These factors maximise the accuracy and reliability of diagnosis using the Olympus true colour LED illuminator.

With a large number of clinical samples to screen, the optimisation of user workflow can significantly improve the efficiency of pathological analysis. For this reason, the Olympus true colour illumination system with LIM automatically modulates light intensity when changing objective lenses. This means that consistent illumination is maintained without the need for manual adjustment, saving a significant amount of time, while simplifying and improving the screening process. Automation is achieved using sensors mounted in the nosepiece that detect the objective lens in use and manipulate the LED intensity according to user defined preferences.

The flexible Olympus BX3 clinical microscopy systems with true colour LED illumination and LIM have been specifically developed to meet the needs of clinical microscopy, ensuring reliable diagnosis while maximising workflow efficiency.

For further information contact, Esther Ahrent, e-mail: microscopy@olympus-europa.com or view website: www.microscopy.olympus.eu   Refer to page 285

Successful North American product launch prompts Saw
Instruments to extend sam®X Early Adopter price promotion

SAW Instruments, developer of the innovative sam® family of biosensors for real-time, label-free assays using living cells, has extended the price promotion surrounding the launch of its new sam®X biosensor. The promotion was so successfully received at the recent North American launch of the product (held at the IBC Antibody Engineering and Antibody Therapeutics meeting in San Diego) that SAW Instruments has decided to make the offer available across Europe and North America. If ordered before the end of 2011, the new system can be obtained at a special early adopter price.

As the high-end member of the sam® family of acoustic biosensors, the new sam®X is designed to offer even greater workflow flexibility and high-throughput capability when analysing molecular interactions on the surface of living mammalian cells. Using Surface Acoustic Wave (SAW) technology, the sam® systems represent an evolution in the way such interactions are measured, producing accurate data much faster than can be achieved using similar equilibrium based approaches.

The new sam®X is designed to offer even greater workflow flexibility and high-throughput capability. when analysing molecular interactions on the surface of living mammalian cells.

Notes on the technology
Surface Acoustic Wave technology is based on the ability of a wave of energy to travel across the surface of a material. Each surface has a typical inherent elasticity affecting the way the energy of the wave dissipates as it travels across the surface of the material being analysed. Thus, the nature of the surface in question, and therefore any changes to it, can be assessed by sensors monitoring the behaviour of the wave as it propagates across the surface. In particular, changes in mass result in alterations to the phase of the wave, whilst viscoelastic and conformational characteristics influence wave amplitude. The technology developed and employed by SAW instruments is capable of accurately interpreting this information in order to provide real time readouts measuring binding and conformational changes in the samples through which the wave passes.

SAW Instruments GmbH in profile
SAW Instruments GmbH designs, develops and sells biosensor-based laboratory instruments for life sciences research. The proprietary technology of SAW Instruments employs a Surface Acoustic Waves approach to biological measurement, rather than using traditional optical methods such as SPR. The technology is uniquely positioned compared to other biosensor technologies based on its ability to measure conformational changes in cells, membranes and vesicles. In addition, the system is also capable of providing other industry standard measurements including those involving kinetics, quantitation and affinity.

SAW instruments has more than ten years experience in the field of label-free biosensor technology and a number of our customers have successfully published work employing the platform. It is our goal is to continue to build on this experience, as well as to develop a family of workflow-driven products to further meet our customers' needs.   

To find out more about the current sam®X price promotion, contact Ian Taylor, e-mail: taylor@saw-instruments.com before the end of the year, or for more information on the sam® family, view website: www.saw-instruments.com   Refer to page 288