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Related Concept Videos

Raman Spectroscopy Instrumentation: Overview01:26

Raman Spectroscopy Instrumentation: Overview

A conventional Raman spectrophotometer includes a laser source, a sample holding system, a wavelength selector, and a detector.
The monochromatic laser source, typically using visible or near-infrared radiation, generates a highly focused beam of light. This light interacts with the molecules of the sample, scattering some of the light. Liquid and gaseous samples are usually tested in ordinary glass capillaries, while solids can be analyzed as powders packed in capillaries or as potassium...
Raman Spectroscopy: Overview01:20

Raman Spectroscopy: Overview

The underlying principle of Raman spectroscopy is based on the interaction between light and matter, specifically molecules' inelastic scattering of photons. When a monochromatic beam of light, typically from a laser source, interacts with a sample, most scattered light has the same frequency as the incident light. This is known as Rayleigh scattering.
However, a small fraction of the scattered light exhibits a frequency shift due to the exchange of energy between the incident photons and the...
Scanning Electron Microscopy01:07

Scanning Electron Microscopy

A scanning electron microscope (SEM) is used to study the surface features of a sample by using an electron beam that scans the sample surface in a two-dimensional manner. Typically, areas between ~1 centimeter to 5 micrometers in width can be imaged. SEM can be used to image bacteria, viruses, tissues as well as larger samples like insects. Conventional SEM gives a magnification ranging from 20X to 30,000X and spatial resolution of 50 to 100 nanometers.
Fundamental Principles
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Related Experiment Video

Updated: Jun 23, 2026

Implementation of a Nonlinear Microscope Based on Stimulated Raman Scattering
09:13

Implementation of a Nonlinear Microscope Based on Stimulated Raman Scattering

Published on: July 6, 2019

Construction of an integrated Raman- and angular-scattering microscope.

Zachary J Smith1, Andrew J Berger

  • 1The Institute of Optics, University of Rochester, Rochester, New York 14627, USA.

The Review of Scientific Instruments
|May 2, 2009
PubMed
Summary

This study introduces a multimodal microscope for analyzing living samples noninvasively. It simultaneously captures elastic and inelastic light scattering, enabling detailed chemical and morphological characterization without dyes.

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Last Updated: Jun 23, 2026

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Area of Science:

  • Biophotonics
  • Microscopy
  • Spectroscopy

Background:

  • Characterizing living biological samples noninvasively is crucial for understanding dynamic biological processes.
  • Existing methods often require exogenous labels or dyes, which can interfere with natural sample evolution.
  • Longitudinal studies of cellular and tissue morphology and chemistry are limited by current imaging techniques.

Purpose of the Study:

  • To develop and demonstrate a multimodal microscope platform for simultaneous elastic and inelastic light scattering measurements.
  • To enable noninvasive, label-free chemical and morphological characterization of living samples.
  • To facilitate longitudinal studies of biological samples over time.

Main Methods:

  • Illumination of a microscopic region (38 µm²) with a 785 nm laser in epi- and transillumination.
  • Simultaneous or sequential collection of elastically and inelastically scattered light.
  • Spectrographic analysis of inelastic scattering for Raman spectra and analysis of elastic scattering for scatterer size distributions.

Main Results:

  • High-quality Raman spectra obtained after fluorescence removal, revealing sample chemical composition.
  • Estimated size distributions of scatterers derived from elastic scattering pupil images using generalized Lorenz-Mie theory.
  • Demonstrated capability for label-free, longitudinal monitoring of living samples.

Conclusions:

  • The developed multimodal microscope platform offers a powerful tool for noninvasive, label-free characterization of biological samples.
  • Simultaneous elastic and inelastic scattering provides complementary information on sample chemistry and morphology.
  • This technique opens new avenues for longitudinal studies of biological systems in their natural state.