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TIRF-SIM (Total internal reflection fluorescence structured illumination microscopy)
About the Innovation
Structured illumination microscopy (SIM) is a method that can double the spatial resolution of wide-field fluorescence microscopy in three dimensions by using spatially structured illumination light. Total internal reflection microscopy increases the signal-to-noise acquired from surface-bound fluorophores by selectively illuminating them.
SIM is fully compatible with the total internal reflection mode, as the researchers from the Betzig laboratory have shown in multiple publications listed here. In TIRF-SIM microscopy, the structured illumination pattern is confined to the TIRF-plane, which is the interface between the glass coverslip and the aqueous medium. Using spatial light modulator, the TIRF-SIM system at Janelia is capable of projecting as well as angle- and phase-shifting the illumination pattern with sufficient speed to support live cell imaging. This allows its users to leverage the gentler SIM illumination for enhanced resolution live cell imaging.
Janelia researchers offer the designs for constructing a TIRF-SIM microscope to take advantage of these features. References to publications and additional information, available via a non-commercial open-source license are available at the Flintbox link. The total information includes background on the theory of SIM and the practical considerations of its optical implementation in different configurations and image reconstruction algorithms. This information is provided for both users of the commercial SIM systems and scientists who wish to develop their own SIM setups.
Free to make for Non-Profit Research by downloading designs at Flintbox (see link at upper right) and rights and designs available for Commercial License.
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