Novel semi-automated fluorescence microscope imaging algorithm for monitoring IgG aggregates in serum

Abstract Analysis of therapeutic IgG aggregates in serum is a potential area of investigation as it can give deeper insights about the function, immunogenic issues and protein interaction associated with the aggregates. To overcome various complexities associated with the existing analytical techniq...

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Autores principales:	Shravan Sreenivasan, Deepak Sonawat, Shyamapada Mandal, Kedar Khare, Anurag S. Rathore
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/233b250b979a49ffa49da7927a7d6bed
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spelling	oai:doaj.org-article:233b250b979a49ffa49da7927a7d6bed2021-12-02T15:02:57ZNovel semi-automated fluorescence microscope imaging algorithm for monitoring IgG aggregates in serum10.1038/s41598-021-90623-72045-2322https://doaj.org/article/233b250b979a49ffa49da7927a7d6bed2021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-90623-7https://doaj.org/toc/2045-2322Abstract Analysis of therapeutic IgG aggregates in serum is a potential area of investigation as it can give deeper insights about the function, immunogenic issues and protein interaction associated with the aggregates. To overcome various complexities associated with the existing analytical techniques for analyzing aggregates in serum, a novel florescence microscopy-based image processing approach was developed. The monoclonal antibody (mAb) was tagged with a fluorescent dye, fluorescein isothiocyanate (FITC). Aggregates, generated by stirring, were spiked into serum and images were captured at various time points. After denoising, thresholding by weighted median, 1D Otsu, and 2D Otsu was attempted and a modified 2D Otsu, a new mode of thresholding, was developed. This thresholding method was found to be highly effective in removing noises and retaining analyte sizes. Out of 0–255, the optimized threshold value obtained for the images discussed in modified 2D Otsu was 9 while 2D Otsu’s overestimated values were 38 and 48. Other morphological operations were applied after thresholding and the area, perimeter, circularity, and radii of the aggregates in these images were calculated. The proposed algorithm offers an approach for analysis of aggregates in serum that is simpler to implement and is complementary to existing approaches.Shravan SreenivasanDeepak SonawatShyamapada MandalKedar KhareAnurag S. RathoreNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-18 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Shravan Sreenivasan Deepak Sonawat Shyamapada Mandal Kedar Khare Anurag S. Rathore Novel semi-automated fluorescence microscope imaging algorithm for monitoring IgG aggregates in serum
description	Abstract Analysis of therapeutic IgG aggregates in serum is a potential area of investigation as it can give deeper insights about the function, immunogenic issues and protein interaction associated with the aggregates. To overcome various complexities associated with the existing analytical techniques for analyzing aggregates in serum, a novel florescence microscopy-based image processing approach was developed. The monoclonal antibody (mAb) was tagged with a fluorescent dye, fluorescein isothiocyanate (FITC). Aggregates, generated by stirring, were spiked into serum and images were captured at various time points. After denoising, thresholding by weighted median, 1D Otsu, and 2D Otsu was attempted and a modified 2D Otsu, a new mode of thresholding, was developed. This thresholding method was found to be highly effective in removing noises and retaining analyte sizes. Out of 0–255, the optimized threshold value obtained for the images discussed in modified 2D Otsu was 9 while 2D Otsu’s overestimated values were 38 and 48. Other morphological operations were applied after thresholding and the area, perimeter, circularity, and radii of the aggregates in these images were calculated. The proposed algorithm offers an approach for analysis of aggregates in serum that is simpler to implement and is complementary to existing approaches.
format	article
author	Shravan Sreenivasan Deepak Sonawat Shyamapada Mandal Kedar Khare Anurag S. Rathore
author_facet	Shravan Sreenivasan Deepak Sonawat Shyamapada Mandal Kedar Khare Anurag S. Rathore
author_sort	Shravan Sreenivasan
title	Novel semi-automated fluorescence microscope imaging algorithm for monitoring IgG aggregates in serum
title_short	Novel semi-automated fluorescence microscope imaging algorithm for monitoring IgG aggregates in serum
title_full	Novel semi-automated fluorescence microscope imaging algorithm for monitoring IgG aggregates in serum
title_fullStr	Novel semi-automated fluorescence microscope imaging algorithm for monitoring IgG aggregates in serum
title_full_unstemmed	Novel semi-automated fluorescence microscope imaging algorithm for monitoring IgG aggregates in serum
title_sort	novel semi-automated fluorescence microscope imaging algorithm for monitoring igg aggregates in serum
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/233b250b979a49ffa49da7927a7d6bed
work_keys_str_mv	AT shravansreenivasan novelsemiautomatedfluorescencemicroscopeimagingalgorithmformonitoringiggaggregatesinserum AT deepaksonawat novelsemiautomatedfluorescencemicroscopeimagingalgorithmformonitoringiggaggregatesinserum AT shyamapadamandal novelsemiautomatedfluorescencemicroscopeimagingalgorithmformonitoringiggaggregatesinserum AT kedarkhare novelsemiautomatedfluorescencemicroscopeimagingalgorithmformonitoringiggaggregatesinserum AT anuragsrathore novelsemiautomatedfluorescencemicroscopeimagingalgorithmformonitoringiggaggregatesinserum
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Novel semi-automated fluorescence microscope imaging algorithm for monitoring IgG aggregates in serum

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