Synthesis of a highly Zn(2+)-selective cyanine-based probe and its use for tracing endogenous zinc ions in cells and organisms.
The zinc ion has a key role in a variety of physiological and pathological processes. As a consequence, the development of sensitive and reliable methods to monitor the presence of zinc ions in cells and organisms is of great importance to biological research and biomedical applications. This protocol describes detailed procedures for the five-stage synthesis of a zinc ion-selective, cyanine-based fluorescent probe, CTMPA, from 2,6-bis(hydroxymethyl)pyridine. In addition, we describe its applications in the detection of Zn(2+) released during apoptosis in cells and endogenous Zn(2+) in living zebrafish. Notably, the use of CTMPA enabled our research group to monitor for the first time the presence of zinc ions in neuromasts of zebrafish via fluorescence. The approximate time frame for the synthesis of CTMPA is 4-5 d, and for its use in bioimaging is 8-10 h for cells and 2 h for zebrafish.
1] Department of Chemistry and Nano Science, Global Top 5 Research Program, Ewha Womans University, Seoul, Korea. [2] Key Laboratory for Advanced Materials and Institute of Fine Chemicals, East China University of Science and Technology, Shanghai, People's Republic of China. [3].
Zhiqian
Z
Guo
1] Center for Biofunctional Molecules, Department of Chemistry, Yonsei University, Seoul, Korea. [2].
Gun-Hee
GH
Kim
Department of Chemistry and Nano Science, Global Top 5 Research Program, Ewha Womans University, Seoul, Korea.
Juyoung
J
Yoon
Center for Biofunctional Molecules, Department of Chemistry, Yonsei University, Seoul, Korea.
Injae
I
Shin
D000818:Animals; D002232:Carbocyanines / Q000737:chemistry; D005453:Fluorescence; D008465:Mechanoreceptors / Q000737:chemistry; D015336:Molecular Probe Techniques; D015335:Molecular Probes / Q000138:chemical synthesis*; D015394:Molecular Structure; D008991:Monitoring, Physiologic / Q000379:methods*; D015027:Zebrafish; D015032:Zinc / Q000032:analysis*
D002232:Carbocyanines; D015335:Molecular Probes; D015032:Zinc