title: Glucagon-Secreting Alpha Cell Selective Two-Photon Fluorescent Probe TP-α: For Live Pancreatic Islet Imaging.
abstract: Two-photon (TP) microscopy has an advantage for live tissue imaging which allows a deeper tissue penetration up to 1 mm comparing to one-photon (OP) microscopy. While there are several OP fluorescence probes in use for pancreatic islet imaging, TP imaging of selective cells in live islet still remains a challenge. Herein, we report the discovery of first TP live pancreatic islet imaging probe; TP-α (Two Photon-alpha) which can selectively stain glucagon secreting alpha cells. Through fluorescent image based screening using three pancreatic cell lines, we discovered TP-α from a TP fluorescent dye library TPG (TP-Green). In vitro fluorescence test showed that TP-α have direct interaction and appear glucagon with a significant fluorescence increase, but not with insulin or other hormones/analytes. Finally, TP-α was successfully applied for 3D imaging of live islets by staining alpha cell directly. The newly developed TP-α can be a practical tool to evaluate and identify live alpha cells in terms of localization, distribution and availability in the intact islets.
authors: Bikram Keshari
authors: BK
authors: Agrawalla
authors: ‡Laboratory of Bio-imaging Probe Development, Singapore Bio-imaging Consortium, Agency of Science, Technology and Research, Singapore 138667, Singapore.
authors: Yogeswari
authors: Y
authors: Chandran
authors: Wut-Hmone
authors: WH
authors: Phue
authors: ‡Laboratory of Bio-imaging Probe Development, Singapore Bio-imaging Consortium, Agency of Science, Technology and Research, Singapore 138667, Singapore.
authors: Sung-Chan
authors: SC
authors: Lee
authors: ‡Laboratory of Bio-imaging Probe Development, Singapore Bio-imaging Consortium, Agency of Science, Technology and Research, Singapore 138667, Singapore.
authors: Yun-Mi
authors: YM
authors: Jeong
authors: ‡Laboratory of Bio-imaging Probe Development, Singapore Bio-imaging Consortium, Agency of Science, Technology and Research, Singapore 138667, Singapore.
authors: Si Yan Diana
authors: SY
authors: Wan
authors: ‡Laboratory of Bio-imaging Probe Development, Singapore Bio-imaging Consortium, Agency of Science, Technology and Research, Singapore 138667, Singapore.
authors: Nam-Young
authors: NY
authors: Kang
authors: ‡Laboratory of Bio-imaging Probe Development, Singapore Bio-imaging Consortium, Agency of Science, Technology and Research, Singapore 138667, Singapore.
authors: Young-Tae
authors: YT
authors: Chang
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D005456:Fluorescent Dyes / Q000737:chemistry*; D050416:Glucagon-Secreting Cells / Q000166:cytology*; D051379:Mice; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D061848:Optical Imaging / Q000379:methods
chemical_list: D005456:Fluorescent Dyes