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

• Published in: STAR protocols Vol. 4; no. 2; p. 102221
• Main Authors: Zhong, Wenying, Oda, Robert, Ozeki, Yasuyuki, Yasui, Masato, Nuriya, Mutsuo
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 16-06-2023; Elsevier
• Subjects: Cell Biology; Cell Culture; Microscopy; Molecular/Chemical Probes; Neuroscience; Protocol; Cell Culture; Neuroscience; Molecular/Chemical Probes; Microscopy; Cell Biology
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2023.102221

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
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•Protocol for real-time cellular imaging using multimodal SRS microscopy•High-resolution imaging of deuterated propofol in living rat neurons•Real-time monitoring of anesthesia dynamics in living neurons•Has potential to be applied to other small-sized drugs
Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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.