Optimised imaging systems for high-speed analysis

1st April 2013

Imaging technologies are now even more optimised to meet the laboratory's main requirements: ease of use and swift analysis of results, Sean Ottewell reports

Innovative imaging technology supplied by Preston-based Image Solutions UK (Imsol) is helping scientists at the University of Sussex's Centre for Genome Damage and Stability (CGDS) to get a better understanding of how mistakes made during the process of cell division can lead to disorders and diseases within humans.

The CGDS houses 16 research groups, all focused on different aspects of DNA damage responses and genome instability. One of the groups, managed by Wellcome Trust senior fellow Helfrid Hochegger, uses Imsol's DeltaVision Core and Personal DV imaging systems in order to better understand the activities of enzymes known as kinases. Successful cell separation depends on kinase activity and as a result they are one of the most studied of all enzyme families.

"My lab is very focused on imaging because we are looking at cell cycle division, especially mitosis. We are particularly interested in the centrosome which organises the dynamics of cell separation.

"Many kinases are involved in this activity and we are trying to understand how they interact by using markers in human and avian tissue culture cells," says Hochegger.

Living tissue

In order to track these markers through the cell, Hochegger and his team have to take 60 images through 300nm-thick slices of living tissue in just 30 seconds. In addition, the images have to be taken with a very short exposure time of just 50mS as any longer would damage the cells that are being studied.

"We film the whole cell and then use deconvolution to reconstruct the image. That's why we need a very sensitive camera and a very stable 3D imaging system. DeltaVision Core and Personal DV are the best solutions because of their control of 3D imaging and ability to optimise it. This is the gold standard for this type of work," he added.

Imsol's DeltaVision Core is designed to increase a laboratory's ability to image more probes and samples over longer periods of time than any competitive imaging system, while the Personal DV is an integrated, bench-top solution designed for speed and economy.

The BioSpectrum imaging system from UVP is configured for use in multiple imaging applications. Combined with the BioLite MultiSpectral light source, it provides a powerful system to excite and illuminate wavelengths for applications including multiplex NIR imaging. The BioSpectrum system provides not only a full range of wavelengths for excitation, also offers rapid, high resolution image capture through the use of cooled CCD camera and low light lenses. Exposures times are very fast, much faster than laser scanning.

The new system features: easy-to-use VisionWorks LS software which automates research with templates for consistent, reproducible results; high resolution and low light sensitivity cooled CCD camera and optics allow quick detection of a full range of visible and NIR fluorescent dyes; automated imaging of white light and multiple different fluorescent tags on a single blot; and a combination of up to eight excitation and five emission wavelengths for all spectral ranges in a single experiment.

Emission filters

With the extensive range of excitation and emission filters available, researchers can detect as well as quantify virtually all fluorescent dye from visible to NIR using VisionWorks LS acquisition and analysis software. The use of NIR and visible fluorescent labels permit multiplexing so that several proteins in a sample can be detected and analysed at the same time on a single protein blot. UVP offers several filters used for NIR blotting with 680 and 770 to 800nm fluorescent tags.

"With the recent release of VisionWorksLS 7.0 software powering the BioSpectrum system, easy and optimal image detection and fast analysis with the simple user interface is now a reality. No matter which fluorophore stain or label is used," says Alex Waluszko, vp for marketing and sales.

NIR imaging

Routine NIR imaging with the BioSpectrum and BioLite MultiSpectral Source is fast, efficient and straightforward yielding full 16-bit images for quantitation and publication. Sean Gallagher, UVP vp and chief technology officer further states, "Automated mulitplex NIR fluorescent imaging with the UVP BioSpectrum further expands capabilities of the most versatile and cost effective fluorescent imaging system available today."

PerkinElmer has debuted an innovative solutions platform for demanding low light level detection applications, called L3D.

The suite of L3D applications solutions contribute to human and environmental health in a range of applications including confocal microscopy, fluorescence and luminescence, molecular/PET imaging, nucleic acid amplification, particle sizing and research and scientific instrumentation.

PerkinElmer's growing range of low light level detection solutions leverages the company's growing range of photon detection technologies and products including avalanche photodiodes (APDs), single photon counting modules (SPCMs), channel photomultipliers (CPMs), and silicon photomultipliers (SiPMs), ideally suited to demanding clinical diagnostics, molecular imaging and analytical instrumentation needs.

Low level light

L3D highlights include: new short wavelength-enhanced Si-APD - PerkinElmer's APDs are ideal for low light level detection applications. Our newest large area short wavelength-enhanced Si-APD is intended for use in demanding molecular imaging and fluorescence detection applications; gigahertz photon detection module (GPDM) - designed for analytical and clinical diagnostic applications under low light level conditions.

The new GPDM delivers the highest dynamic range available in a single operating mode and very low noise; single photon counting modules (SPCMs) - the company says its cutting-edge SPCMs deliver high photon detection efficiencies (PDE) and very low dark noise for confocal microscopy, particle sizing and time-correlated single photon counting (TCSPC) applications; and silicon photomultipliers (SiPMs) - PerkinElmer recently entered into an exclusive agreement with Max Planck Innovation for the licensing of its ultra-fast, low cross-talk SiPM technology. The technology offers record-setting PDE and ultra-short response time. SiPMs offer OEMs a solid state alternative to traditional PMTs in terms of sensitivity and cost.

High content screening

Meanwhile, Molecular Devices' ImageXpress Micro Widefield High Content Screening System is said to offer the ultimate combination of speed and flexibility in a turnkey solution that streamlines research and drug discovery in a fully integrated high content screening environment.

Built on over 10 years of automated imaging experience, the ImageXpress Micro System captures research-quality images with what the company says is the widest range (1-100x) of objective lenses available in an HCS system, enabling users to work at the resolution appropriate for their biology, including small organism, cellular events, or intracellular events. The system's best-in-class 100 nm resolution stages precisely locate and identify sub-cellular features, ensuring excellent X, Y, and Z positioning to simplify tracing individual cells during multi-day time-lapse experiments or mapping neuronal outgrowth across multiple tiled images.

Cellular analysis is made easy with the setup wizard that allows any user in the laboratory to acquire and process preconfigured experiments through an intuitive interface.

Industry-leading stage and focus control, coupled with a high-speed laser autofocus option, make the ImageXpress Micro System the fastest, most precise automated CCD imaging station available.

Combined with mature software, the custom designed ImageXpress Micro Instrument outperforms automated microscopes built from off-the-shelf components, providing a robust high content screening (HCS) platform to address both research and drug discovery applications.






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