Abstract:
:In principle, the long emission lifetimes of lanthanide chelates should enable their ultrasensitive detection in biological systems by time-resolved optical microscopy. However, most lanthanide-imaging systems cannot achieve sensitivities that exceed those of conventional fluorescence microscopes, since they are limited by inefficient lanthanide excitation, the low photon flux of excited lanthanide luminophores, and optics-derived background photoluminescence. We recently reported a new lanthanide-imaging modality, trans-reflected illumination with luminescence resonance energy transfer (trLRET), which overcomes each of these constraints. Here we provide a detailed procedure for visualizing endogenous protein expression in zebrafish embryos, using lanthanide-labeled antibodies, Q-switched laser illumination, and trLRET microscopy. These methods allow ultrasensitive molecular imaging in cells and organisms, establishing a new paradigm for biological exploration and discovery.
journal_name
Methods Enzymoljournal_title
Methods in enzymologyauthors
Ciepla P,Cho U,Chen JKdoi
10.1016/bs.mie.2020.04.030subject
Has Abstractpub_date
2020-01-01 00:00:00pages
225-248eissn
0076-6879issn
1557-7988pii
S0076-6879(20)30158-0journal_volume
640pub_type
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