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In recent years there has been a growing trend to work with live cells, for instance, in high-throughput screening for drug discovery, for stem cell research or for such applications as in vitro fertilization.

Tanuja Koppal, PhD

Advances in Optics Design and the Availability of Integrated Systems Drive new Applications for Cellular Microscopy

In recent years there has been a growing trend to work with live cells, for instance, in high-throughput screening for drug discovery, for stem cell research or for such applications as in vitro fertilization. “Certainly there is a lot more interest in live-cell imaging for looking at dynamic events, and microscopes are being built to achieve that,” says Joseph LoBiondo, product planning manager for Nikon Instruments Inc. “There is a lot of optics design going into achieving live-cell imaging.”

Self-contained, fully integrated live-cell imaging systems now come equipped with a built-in cell incubation chamber and a microscopy unit. Such companies as PerkinElmer, Carl Zeiss, Molecular Devices and Nikon all offer imaging systems with environmental controls for temperature, CO2 levels and humidity in order to ensure that cells can grow and survive for an extended period of time. “Cells stay alive for days in these incubation systems,” says LoBiondo. And in some instruments the cells don’t ever have to be taken out of the system for observation since the microscope is a part of the controlled environment. Units also come equipped with a full-sized incubator that can hold a variety of chamber slides and well plates. “Nikon’s BioStation CT has a motorized arm that pulls out the specified dish or plate, with minimum vibration or disruption to the cells,” says LoBiondo. “The unit is very methodical and slow and very carefully picks up the dish and takes it to the microscope.” The units also have some level of built-in security provided. “Certain program locks can be put in place so that if [researchers] are doing different experiments, each researcher will have access to only [his or her] cells.”

Manufacturers started offering integrated units when interest in live-cell imaging began to grow. Initially researchers, themselves, were integrating such individual components as motorized stages, filter wheels, shutters, cameras and software packages from a number of different manufacturers to create a system that would meet their needs. “Depending on how it is done, there are a lot of components that are needed to work together in order to make an integrated system,” says LoBiondo. “Integrated systems also tend to be less expensive. It could be about half to one-third of the costs of buying and putting together individual components.” The disadvantage is that an integrated system is less customizable and may compromise certain features, such as speed when switching between wavelengths, or flexibility in terms of number of objectives or software that can be used (the latter being crucial for certain applications). However, integrated systems work well for those laboratories in need of multiple units that can perform in a routine and reliable fashion but lack the time or expertise to build them.

However, with microscopes now being designed to cater to several different applications and supported by image analysis software and high-end computing hardware systems, matching products to a specific application may soon be a thing of the past. “Microscopes now have multimode capabilities to be used in multiuse facilities,” says LoBiondo. “They can now switch between total internal reflection fluorescence, confocal and live-cell imaging, with software control and motorization to do all those modalities.”

Soon we may have a system that can do it all.

Tanuja Koppal, PhD, is a freelance science writer and consultant based in Randolph, N.J.

The iVue and iScope allow precision optical analysis and image-sharing with plug-and-go simplicity. Each instrument is an advanced video inspection system, while the iScope enlarges from 10X to 44X and has screen magnification up to 166X. By combining a 3-megapixel camera and touch-screen virtual keyboard for annotation and measurement, the self-contained systems eliminate compatibility concerns, reduce clutter and ease portability. No laptop or desktop PC is needed, unlike other video microscope systems. The iVue’s tilting head provides not only a top-down view, but also an angled view for increased depth of field. The iScope has a 45-degree fixed 3-D viewer option that rotates for 360-degree views without moving the object being examined. Each offers numerous measurement options in various orientations without requiring additional software. Precise distances between points or geometries are calculated on-screen and saved when an image is captured.

The Quanta™ 50 Series is a multipurpose scanning electron microscope (SEM) for labs that require highperformance imaging for a broad range of samples, including those that are insulating, wet, dirty, or dynamically changing, in industries such as materials research, mineralogy, chemicals and petroleum, electronics, pharmaceuticals and biology. Some technologies used in the Magellan™ Extreme High Resolution SEM (XHR SEM) have been adapted to the Quanta system. Specifically, beam deceleration increases the surface imaging capability with lower landing energies, and SmartSCAN™ technologies further improve image quality by reducing noise. This adds high-vacuum, low-landing energy imaging capability to the flexibility of the Quanta’s low-vacuum and environmental SEM (ESEM) technology. ESEM, pioneered by FEI, enables the broadest range of in situ imaging conditions.

The McCrone Group
The Olympus SZX10 stereo-microscope with coaxial illumination module and PAXcam digital camera helps users maximize information available from their samples with a 600 line pairs per millimeter resolution and zoom ratio of 10:1, making analysis more efficient and precise. With the common objective design it is a fully modular system offering a range of options. The coaxial illumination module then enhances contrast, increasing visual detail well beyond the ability of other stereomicroscopes. While the PAXcam digital camera equipped with Microsoft compatible measurement software precisely captures images and analyzes them with grain sizing, particle counting and image stitching. Users are able to fully document their experiments and samples and easily place them into their existing software systems.

Carl Zeiss Microimaging
The LSM 7 MP laser scanning microscope generates high resolution images for a wide range of experiments in biomedical and basic research. Thanks to the use of two scanners in one compact system, two lasers with different wavelengths can be utilized sequentially or simultaneously for specimen imaging and manipulation. The range of detectors and their filter equipment, as well as an extensive line of microscope accessories, allow customization of the microscope system. The system is designed for recording high resolution images of fluorescent structures. The application fields extend from high resolution 3D imaging in long-term observations of development processes and functional imaging in conjunction with simultaneous photo-manipulation right up to imaging in combination with electrophysiological measuring methods.

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