DeltaVision|OMXTM platforms are based on the OMX microscopy system developed by scientists at the University of California, San Francisco (UCSF) and exclusively licensed to Applied Precision. DeltaVision|OMX consists of a newly designed optical platform that greatly improves light throughput, alignment and thermal stability compared to traditional microscope stands. These improvements were an essential development to enable super resolution imaging using 3D structured illumination (3D-SIM) technology invented by the UCSF team led by Drs. Sedat, Agard, and Gustafsson.
Applied Precision has led the way over the past fifteen years empowering scientists to surpass microscope resolution limits through image restoration (deconvolution) microscopy. Our DeltaVision Core and personalDV Microscopy Systems improve resolution by approx 15-20% and contrast by ten fold. In recent years a variety of new approaches have been developed to surpass the diffraction limit. Collectively referred to as Super Resolution Microscopy, these methods allow precise visualization and measurement of features that are less than one-half of the size of those seen with conventional microscopy.
Ultimately the resolution of a microscope system is physically limited by the numerical aperture of the objective lens. However by employing other approaches such as Structured Illumination, Photoactivable Fluorophores (PALM and STORM), TIRF imaging and STED imaging, it is possible to image objects beyond this diffraction limit.
Structured illumination, as utilized exclusively by the DeltaVision|OMX systems, is the only approach which gives true 3D super resolution and can be employed on standard samples without the requirement for special fluorophores or genetically engineered markers.
Comparison of a DeltaVision OMX super resolution image and a immunogold TEM micrograph.
(PCNA localization in CHO cell nuclei. Used with permission from Andrew Belmont,
University of Illinois, Urbana-Champaign.)
Three Dimensional Structured Illumination Microscopy (3D-SIM) works by projecting a structured light pattern onto the sample. The illumination pattern interacts with the fluorescent probes in the sample to generate a new pattern (moiré). By modulating the illumination pattern and interpreting the subsequent images, the reconstruction software is able to determine the distribution of the probes in sample to effectively twice the diffraction limit.
Three dimensional structured illumination microscopy goes a step further than other structured light systems by superimposing a 3 dimensional light pattern onto the sample. The result is that super resolution is achieved in X, Y and Z dimensions simultaneously. The effectiveness is limited only by how deep into the sample the illumination pattern can be maintained before light scattering and optical distortions degrade the contrast (usually tens of microns into the sample).
In addition to the above, the 3D-SIM pattern can be efficiently generated with multiple excitation wavelengths. This allows use of multiple fluorophores in the same sample. Due to the design of the OMX optical platform it is possible to simultaneously image multiple probes in the same sample (currently up to 4) in three dimensional super resolution.
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