Two-Photon Dye Cocktail for Dual-Color 3D Imaging of Pancreatic Beta and Alpha Cells in Live Islets.
Insulin-secreting beta cells together with glucagon-producing alpha cells play an essential role in maintaining the optimal blood glucose level in the body, so the development of selective probes for imaging of these cell types in live islets is highly desired. Herein we report the development of a 2-glucosamine-based two-photon fluorescent probe, TP-β, that is suitable for imaging of beta cells in live pancreatic islets from mice. Flow cytometry studies confirmed that TP-β is suitable for isolation of primary beta cells. Moreover, two-photon imaging of TP-β-stained pancreatic islets showed brightly stained beta cells in live islets. Insulin enzyme-linked immunosorbent assays revealed that TP-β has no effect on glucose-stimulated insulin secretion from the stained islet. Finally, to develop a more convenient islet imaging application, we combined our recently published alpha-cell-selective probe TP-α with TP-β to make a "TP islet cocktail". This unique dye cocktail enabled single excitation (820 nm) and simultaneous dual-color imaging of alpha cells (green) and beta cells (red) in live pancreatic islets. This robust TP islet cocktail may serve as a valuable tool for basic diabetic studies.
Department of Chemistry and Medicinal Chemistry Program, National University of Singapore , 3 Science Drive 3, 117543 Singapore.
Bikram Keshari
BK
Agrawalla
Department of Energy Systems Research, Ajou University , Suwon 443749, Korea.
Hyo Won
HW
Lee
Department of Chemistry and Medicinal Chemistry Program, National University of Singapore , 3 Science Drive 3, 117543 Singapore.
Wut-Hmone
WH
Phue
Laboratory of Bioimaging Probe Development, Singapore Bioimaging Consortium, Agency for Science, Technology and Research , 11 Biopolis Way, # 02-02 Helios, 138667 Singapore.
Anandhkumar
A
Raju
Department of Chemistry and Medicinal Chemistry Program, National University of Singapore , 3 Science Drive 3, 117543 Singapore.
Jong-Jin
JJ
Kim
Department of Energy Systems Research, Ajou University , Suwon 443749, Korea.
Hwan Myung
HM
Kim
Laboratory of Bioimaging Probe Development, Singapore Bioimaging Consortium, Agency for Science, Technology and Research , 11 Biopolis Way, # 02-02 Helios, 138667 Singapore.
Nam-Young
NY
Kang
Department of Chemistry and Medicinal Chemistry Program, National University of Singapore , 3 Science Drive 3, 117543 Singapore.|Laboratory of Bioimaging Probe Development, Singapore Bioimaging Consortium, Agency for Science, Technology and Research , 11 Biopolis Way, # 02-02 Helios, 138667 Singapore.
Young-Tae
0000-0002-1927-3688
YT
Chang
D002470:Cell Survival; D002478:Cells, Cultured; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D005944:Glucosamine / Q000138:chemical synthesis / Q000737:chemistry*; D006801:Humans; D021621:Imaging, Three-Dimensional; D050417:Insulin-Secreting Cells / Q000166:cytology*; D015394:Molecular Structure; D017785:Photons
D005456:Fluorescent Dyes; D005944:Glucosamine