Please use this identifier to cite or link to this item: https://libjncir.jncasr.ac.in/xmlui/handle/10572/1998
Title: Multispectral Bayesian reconstruction technique for real-time two color fluorescence microscopy
Authors: Dilipkumar, Shilpa
Manjithaya, Ravi
Mondal, Partha Pratim
Keywords: Chemistry
Structured Illumination
Disaccharide Moiety
Protein
Aggregation
Huntingtin
Bleomycin
Autophagy
Cells
Issue Date: 2015
Publisher: Royal Society of Chemistry
Citation: RSC Advances
5
17
Dilipkumar, S.; Manjithaya, R.; Mondal, P. P., Multispectral Bayesian reconstruction technique for real-time two color fluorescence microscopy. RSC Advances 2015, 5 (17), 13175-13183.
Abstract: We have developed a real-time imaging method for two-color wide-field fluorescence microscopy using a combined approach that integrates multi-spectral imaging and Bayesian image reconstruction technique. To enable simultaneous observation of two dyes (primary and secondary), we exploit their spectral properties that allow parallel recording in both the channels. The key advantage of this technique is the use of a single wavelength of light to excite both the primary dye and the secondary dye. The primary and secondary dyes respectively give rise to fluorescence and bleed-through signal, which after normalization were merged to obtain two-color 3D images. To realize real-time imaging, we employed maximum likelihood (ML) and maximum a posteriori (MAP) techniques on a high-performance computing platform (GPU). The results show two-fold improvement in contrast while the signal-to-background ratio (SBR) is improved by a factor of 4. We report a speed boost of 52 and 350 for 2D and 3D images respectively. Using this system, we have studied the real-time protein aggregation in yeast cells and HeLa cells that exhibits dot-like protein distribution. The proposed technique has the ability to temporally resolve rapidly occurring biological events.
Description: Restricted access
URI: https://libjncir.jncasr.ac.in/xmlui/10572/1998
ISSN: 2046-2069
Appears in Collections:Research Papers (Ravi Manjithaya)

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