Multicolor Live-Cell Chemical Imaging by Isotopically Edited Alkyne Vibrational Palette

详细信息    查看全文

• 作者：Zhixing Chen ; Daniel W. Paley ; Lu Wei ; Andrew L. Weisman ; Richard A. Friesner ; Colin Nuckolls ; Wei Min
• 刊名：Journal of the American Chemical Society
• 出版年：2014
• 出版时间：June 4, 2014
• 年：2014
• 卷：136
• 期：22
• 页码：8027-8033
• 全文大小：414K
• 年卷期：v.136,no.22(June 4, 2014)
• ISSN：1520-5126

文摘

Vibrational imaging such as Raman microscopy is a powerful technique for visualizing a variety of molecules in live cells and tissues with chemical contrast. Going beyond the conventional label-free modality, recent advance of coupling alkyne vibrational tags with stimulated Raman scattering microscopy paves the way for imaging a wide spectrum of alkyne-labeled small biomolecules with superb sensitivity, specificity, resolution, biocompatibility, and minimal perturbation. Unfortunately, the currently available alkyne tag only processes a single vibrational 鈥渃olor鈥? which prohibits multiplex chemical imaging of small molecules in a way that is being routinely practiced in fluorescence microscopy. Herein we develop a three-color vibrational palette of alkyne tags using a ¹³C-based isotopic editing strategy. We first synthesized ¹³C isotopologues of EdU, a DNA metabolic reporter, by using the newly developed alkyne cross-metathesis reaction. Consistent with theoretical predictions, the mono-¹³C (¹³C鈮?sup>12C) and bis-¹³C (¹³C鈮?sup>13C) labeled alkyne isotopologues display Raman peaks that are red-shifted and spectrally resolved from the originally unlabeled (¹²C鈮?sup>12C) alkynyl probe. We further demonstrated three-color chemical imaging of nascent DNA, RNA, and newly uptaken fatty-acid in live mammalian cells with a simultaneous treatment of three different isotopically edited alkynyl metabolic reporters. The alkyne vibrational palette presented here thus opens up multicolor imaging of small biomolecules, enlightening a new dimension of chemical imaging.