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• W2309494507 abstract "Abstract Secretory granules (SGs) in mast cells contain various molecules that elicit allergy symptoms and are generally considered therapeutic targets. However, the biogenesis, maintenance, regulation and recycling of these granules remain controversial, mainly due to the lack of suitable live-cell imaging methods. In this study, we applied negative contrast imaging with soluble green fluorescent protein (GFP) expressed in the cytoplasm as a method to validate structural information of mast cell SGs. We evaluated the accuracy of the method in detail and we demonstrated that it can be used for quantitative analysis. Using this technique, secretory granules, the nucleus, mitochondria and the cell body were visualized in individual RBL-2H3 mast cells without any influence. When combined with conventional multicolor fluorescence imaging, visualization of SG-associated proteins and SG–SG fusion was achieved. Moreover, 3D images were constructed based on this method and detailed information on the number, size and shape of individual SGs was obtained. We found that cell volume was correlated with SG number. In summary, the technique provides valuable and unique data and will therefore advance SG research." @default.
• W2309494507 created "2016-06-24" @default.
• W2309494507 creator A5002377181 @default.
• W2309494507 creator A5010738368 @default.
• W2309494507 date "2016-03-21" @default.
• W2309494507 modified "2023-10-16" @default.
• W2309494507 title "A method for detailed analysis of the structure of mast cell secretory granules by negative contrast imaging" @default.
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• W2309494507 doi "https://doi.org/10.1038/srep23369" @default.
• W2309494507 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/4800307" @default.
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