Abstract:
:Resin embedding is widely used and facilitates microscopic imaging of biological tissues. In contrast, quenching of fluorescence during embedding process hinders the application of resin embedding for imaging of fluorescence-labeled samples. For samples expressing fluorescent proteins, it has been demonstrated that the weakened fluorescence could be recovered by reactivating the fluorophore with alkaline buffer. We extended this idea to immunofluorescence-labeling technology. We showed that the fluorescence of pH-sensitive fluorescein isothiocyanate (FITC) was quenched after resin embedding but reactivated after treating by alkaline buffer. We observed 138.5% fluorescence preservation ratio of reactivated state, sixfold compared with the quenched state in embedding resin, which indicated its application for fluorescence imaging of high signal-to-background ratio. Furthermore, we analyzed the chemical reactivation mechanism of FITC fluorophore. This work would show a way for high-resolution imaging of immunofluorescence-labeled samples embedded in resin.
journal_name
J Biomed Optjournal_title
Journal of biomedical opticsauthors
Li L,Rao G,Lv X,Chen R,Cheng X,Wang X,Zeng S,Liu Xdoi
10.1117/1.JBO.23.2.020501subject
Has Abstractpub_date
2018-02-01 00:00:00pages
1-4issue
2eissn
1083-3668issn
1560-2281pii
JBO-170700LRjournal_volume
23pub_type
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