Fluoromodule-based reporter/probes designed for in vivo fluorescence imaging.

The Journal of Clinical Investigation
Ming ZhangAlan S Waggoner

Abstract

Optical imaging of whole, living animals has proven to be a powerful tool in multiple areas of preclinical research and has allowed noninvasive monitoring of immune responses, tumor and pathogen growth, and treatment responses in longitudinal studies. However, fluorescence-based studies in animals are challenging because tissue absorbs and autofluoresces strongly in the visible light spectrum. These optical properties drive development and use of fluorescent labels that absorb and emit at longer wavelengths. Here, we present a far-red absorbing fluoromodule-based reporter/probe system and show that this system can be used for imaging in living mice. The probe we developed is a fluorogenic dye called SC1 that is dark in solution but highly fluorescent when bound to its cognate reporter, Mars1. The reporter/probe complex, or fluoromodule, produced peak emission near 730 nm. Mars1 was able to bind a variety of structurally similar probes that differ in color and membrane permeability. We demonstrated that a tool kit of multiple probes can be used to label extracellular and intracellular reporter-tagged receptor pools with 2 colors. Imaging studies may benefit from this far-red excited reporter/probe system, which features tight co...Continue Reading

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Citations

Dec 19, 2015·BMC Biology·Joshua C SnyderMarc G Caron
Apr 4, 2016·Journal of Controlled Release : Official Journal of the Controlled Release Society·Wen LiuWilson S Meng
Apr 28, 2016·ACS Chemical Biology·Larry S BarakMarc G Caron
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Mar 25, 2017·Oncology Reports·Zongqiang LaiYongxiang Zhao
Jan 13, 2019·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Erika J CrosbyZachary C Hartman
Jun 25, 2021·Chemical Science·Shahi Imam RejaKazuya Kikuchi
May 10, 2016·Bioconjugate Chemistry·Matharishwan NaganbabuMarcel P Bruchez
Aug 19, 2017·Organic Letters·Qingyang ZhangHai-Yu Hu

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Methods Mentioned

BETA
fluorescence imaging
flow cytometry
confocal microscopy
PCR
column chromatography

Software Mentioned

Andor IQ
SPSS
TrueQuant
ImageJ
QuickCal
FlowJo X

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