Protocol to image deuterated propofol in living rat neurons using multimodal stimulated Raman scattering microscopy

Wenying Zhong; Robert Oda; Yasuyuki Ozeki; Masato Yasui; Mutsuo Nuriya

doi:10.1016/j.xpro.2023.102221

Protocol to image deuterated propofol in living rat neurons using multimodal stimulated Raman scattering microscopy

STAR Protoc. 2023 Apr 13;4(2):102221. doi: 10.1016/j.xpro.2023.102221. Online ahead of print.

Authors

Wenying Zhong¹, Robert Oda², Yasuyuki Ozeki³, Masato Yasui¹, Mutsuo Nuriya⁴

Affiliations

¹ Department of Pharmacology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku, Tokyo 160-8582, Japan.
² Department of Pharmacology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku, Tokyo 160-8582, Japan; Department of Electrical Engineering and Information Systems, Graduate School of Engineering, 7-3-1 Hongo, Bunkyo, Tokyo 113-8656, Japan.
³ Department of Electrical Engineering and Information Systems, Graduate School of Engineering, 7-3-1 Hongo, Bunkyo, Tokyo 113-8656, Japan.
⁴ Department of Pharmacology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku, Tokyo 160-8582, Japan; Graduate School of Environment and Information Sciences, Yokohama National University, 79-1 Tokiwadai, Hodogaya, Yokohama, Kanagawa 240-8501, Japan. Electronic address: mnuriya@keio.jp.

Abstract

Propofol is a widely used anesthetic important in clinics, but like many other bioactive molecules, it is too small to be tagged and visualized by fluorescent dyes. Here, we present a protocol to visualize deuterated propofol in living rat neurons using stimulated Raman scattering (SRS) microscopy with carbon-deuterium bonds serving as a Raman tag. We describe the preparation and culture of rat neurons, followed by optimization of the SRS system. We then detail neuron loading and real-time imaging of anesthesia dynamics. For complete details on the use and execution of this protocol, please refer to Oda et al.¹.

Keywords: Cell Biology; Cell Culture; Microscopy; Molecular/Chemical Probes; Neuroscience.