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BIOIMAGING IS CURRENTLY UNDERGOING AN EXCITING revolution. This includes all aspects of imaging from probe development, specimen preparation, and instrumentation to image analysis and quantitation. Perhaps the most exciting developments are new platforms for imag- ing that radically advance the capabilities for collecting high-spatial and high-temporal resolu- tion data. In many cases, the standard microscope has been replaced with new purpose-built plat- forms that are much more flexible and enable the implementation of new imaging modalities such as particular single-molecule and super-resolution imaging methods. For live cell imaging, there are a number of competing critical requirements. Any live cell imag- ing system must be physically stable so that vibrations and temperature shifts do not move the sam- ple or the optical path. This requirement is undermined by the need to change focus and to collect images as rapidly as possible. Fast live cell imaging thus requires a very stable, rapidly, and accurately moving imaging system with little vibration or temperature change. Photobleaching and photo- damage limit the photon budget, the number of photons transmitted through the microscope. When working at photon-limited levels, any additional sources of background and noise, such as stray light or noise from camera electronics, must be avoided. A recent study has highlighted the presence of additional non-Poisson noise in all tested commercial imaging platforms (Murray et al. 2007). Thus, photon-limited imaging is extremely challenging on traditional microscope platforms. In this chapter, we discuss the design principles and applications of the OMX microscope, a new platform that provides unprecedented mechanical and thermal stabilities coupled with a photon budget that is dramatically improved over traditional microscope platforms. These char- acteristics make the OMX microscope outstanding for fast live cell imaging and super-resolution imaging. Moreover, its open flexible architecture makes it particularly amenable to adding other modes of microscopy to the platform.

Type

Journal article

Publisher

Cold Spring Harbor

Publication Date

09/2009

Pages

1 - 12