Raman Spectroscopy, Microscopy and Imaging Q&A Booklet

   
Raman spectroscopy is well established as one of the most powerful and versatile techniques for a diverse range of applications in both research and analytical laboratories. Integrating the Raman spectrometer with a light microscope offers a unique ability to noninvasively characterize chemically complex and spatially inhomogeneous samples with a sub-micron spatial resolution. Modern confocal Raman microscopy and imaging, which allows one to obtain two- and three-dimensional spectrochemical images of samples in various states and forms, has become an indispensable research method for scientific communities, from physicists to chemists, to criminalists, geologists, biologists, medical professionals, and others.

The Raman Spectroscopy, Microscopy and Imaging Q&A Booklet aims to familiarize scientists across a broad range of disciplines with the fundamentals of modern Raman spectroscopy and microscopy in particular.
  

The first section outlines the physics of the interaction between light and molecules and the experimental discovery of the Raman effect.

The second section introduces the basics of Raman scattering and Raman spectroscopy. It considers the relationship between the properties of molecules and their Raman activity, points out the complementary nature of Raman and FTIR spectroscopies, and summarizes the major benefits of Raman spectroscopy. Special attention is drawn to the phenomenon of fluorescence and how it interferes with Raman spectra.

In the third section, the key components of a Raman spectrometer are specified, the basic principles of their operations are described, and their main parameters are characterized. Emphasis is placed on the importance of choosing the most optimal excitation wavelength of laser light and finely controlling its power in order to obtain the most informative Raman spectral data for a specific application.
  
The final fourth section introduces the fundamentals and practical aspects of confocal Raman microscopy. The role of confocal aperture, the importance of regular alignment and calibration procedures, and essential relations between the microscope objective, aperture setting and mapping step are emphasized.  The recent advances in fast Raman imaging technique, capable of obtaining both traditional optical and spectrochemical images with a spatial resolution at the diffraction limit of light, are outlined. The section is oriented towards helping readers understand and successfully operate a Raman microscope.

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