17 February 2017 Spatially-controlled illumination with rescan confocal microscopy enhances image quality, resolution and reduces photodamage
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Abstract
Fluorescence microscopy is an important tool in biomedical imaging. An inherent trade-off lies between image quality and photodamage. Recently, we have introduced rescan confocal microscopy (RCM) that improves the lateral resolution of a confocal microscope down to 170 nm. Previously, we have demonstrated that with controlled-light exposure microscopy, spatial control of illumination reduces photodamage without compromising image quality. Here, we show that the combination of these two techniques leads to high resolution imaging with reduced photodamage without compromising image quality. Implementation of spatially-controlled illumination was carried out in RCM using a line scanning-based approach. Illumination is spatially-controlled for every line during imaging with the help of a prediction algorithm that estimates the spatial profile of the fluorescent specimen. The estimation is based on the information available from previously acquired line images. As a proof-of-principle, we show images of N1E-115 neuroblastoma cells, obtained by this new setup with reduced illumination dose, improved resolution and without compromising image quality.
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Venkataraman Krishnaswami, Venkataraman Krishnaswami, Giulia M. R. De Luca, Giulia M. R. De Luca, Ronald M. P. Breedijk, Ronald M. P. Breedijk, Cornelis J. F. Van Noorden, Cornelis J. F. Van Noorden, Erik M. M. Manders, Erik M. M. Manders, Ron A. Hoebe, Ron A. Hoebe, } "Spatially-controlled illumination with rescan confocal microscopy enhances image quality, resolution and reduces photodamage", Proc. SPIE 10070, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIV, 100700G (17 February 2017); doi: 10.1117/12.2251329; https://doi.org/10.1117/12.2251329
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