Improving signal-to-noise ratio of structured light microscopy based on photon reassignment

In this paper, we report a method for 3D visualization of a biological specimen utilizing a structured light wide-field microscopic imaging system. This method improves on existing structured light imaging modalities by reassigning fluorescence photons generated from off-focal plane excitation, impr...

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Bibliographic Details
Main Authors: Singh, Vijay Raj, Choi, Heejin, Yew, Elijah Y. S., Bhattacharya, Dipanjan, Yuan, Luo, Sheppard, Colin J. R., Rajapakse, Jagath C., Barbastathis, George, So, Peter T. C.
Other Authors: School of Computer Engineering
Format: Article
Language:English
Published: 2013
Online Access:https://hdl.handle.net/10356/96324
http://hdl.handle.net/10220/10165
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Institution: Nanyang Technological University
Language: English
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Summary:In this paper, we report a method for 3D visualization of a biological specimen utilizing a structured light wide-field microscopic imaging system. This method improves on existing structured light imaging modalities by reassigning fluorescence photons generated from off-focal plane excitation, improving in-focus signal strength. Utilizing a maximum likelihood approach, we identify the most likely fluorophore distribution in 3D that will produce the observed image stacks under structured and uniform illumination using an iterative maximization algorithm. Our results show the optical sectioning capability of tissue specimens while mostly preserving image stack photon count, which is usually not achievable with other existing structured light imaging methods.