Microsecond scale vibrational spectroscopic imaging by multiplex stimulated Raman scattering microscopy

Chien-Sheng Liao; Mikhail N Slipchenko; Ping Wang; Junjie Li; Seung-Young Lee; Robert A Oglesbee; Ji-Xin Cheng

doi:10.1038/lsa.2015.38

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Microsecond scale vibrational spectroscopic imaging by multiplex stimulated Raman scattering microscopy

Original Articles

- Microsecond scale vibrational spectroscopic imaging by multiplex stimulated Raman scattering microscopy
- Light: Science & Applications Vol. 4, Issue 2, (2015)
- Affiliations：
  
  1.Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN 47907, USA
  2.Department of Biological Sciences, Purdue University, West Lafayette, IN 47907, USA
  3.Department of Chemistry, Purdue University, West Lafayette, IN 47907, USA
- Author bio：
  
  JX Cheng, Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN 47907, USA E-mail: jcheng@purdue.edu
- Funds：
- DOI：10.1038/lsa.2015.38
  CLC：
- Published：2015-06，
  
  Published Online：13 January 2015，
  
  Received：05 August 2014，
  
  Revised：09 January 2015，
  
  Accepted：10 January 2015
- Accepted：
Scan QR Code
Liao, C. S. et al. Microsecond scale vibrational spectroscopic imaging by multiplex stimulated Raman scattering microscopy. Light: Science & Applications, 4, e265 (2015).
DOI：

Liao, C. S. et al. Microsecond scale vibrational spectroscopic imaging by multiplex stimulated Raman scattering microscopy. Light: Science & Applications, 4, e265 (2015). DOI： 10.1038/lsa.2015.38.

摘要

Abstract

Real-time vibrational spectroscopic imaging is desired for monitoring cellular states and cellular processes in a label-free manner. Raman spectroscopic imaging of highly dynamic systems is inhibited by relatively slow spectral acquisition on millisecond to second scale. Here

we report microsecond scale vibrational spectroscopic imaging by lock-in free parallel detection of spectrally dispersed stimulated Raman scattering signal. Using a homebuilt tuned amplifier array

our method enables Raman spectral acquisition

within the window defined by the broadband pulse

at the speed of 32 µs and with close to shot-noise limited detection sensitivity. Incorporated with multivariate curve resolution analysis

our platform allows compositional mapping of lipid droplets in single live cells

observation of intracellular retinoid metabolism

discrimination of fat droplets from protein-rich organelles in

Caenorhabditis elegans

spectral detection of fast flowing tumor cells and monitoring drug diffusion through skin tissue

in vivo

. The reported technique opens new opportunities for compositional analysis of cellular compa

rtment in a microscope setting and high-throughput spectral profiling of single cells in a flow cytometer setting.

关键词

Keywords

optical microscopyRaman scatteringvibrational spectroscopy

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