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Related Concept Videos

Cryo-electron Microscopy01:28

Cryo-electron Microscopy

Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...
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...

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Cryo-Structured Illumination Microscopic Data Collection from Cryogenically Preserved Cells
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The feasibility of Cryo In-SEM Raman microspectroscopy.

J Hazekamp1, M G Reed, C V Howard

  • 1Unilever R&D Vlaardingen, The Netherlands. Johan.Hazekamp@unilever.com

Journal of Microscopy
|June 24, 2011
PubMed
Summary
This summary is machine-generated.

Cryogenic preparation methods preserve sample chemistry and ultrastructure for combined Raman microspectrometry and scanning electron microscopy (SEM). This technique enhances spectral band sharpening, improving detailed analysis of specimens.

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Area of Science:

  • Materials Science
  • Analytical Chemistry
  • Microscopy

Background:

  • Standard scanning electron microscopy (SEM) specimen preparation introduces chemical and structural artifacts, compromising accurate compositional analysis.
  • Cryo preparation methods are essential for preserving the native chemical composition and ultrastructure of samples.
  • Combining Raman microspectrometry with SEM offers advanced analytical capabilities, but requires compatible specimen handling.

Purpose of the Study:

  • To develop and implement a cryo transfer system for combined Raman microspectrometry and SEM (EMRAM instrument).
  • To evaluate the impact of cryogenic temperatures on Raman spectra of model specimens.
  • To assess the preservation of chemical composition and ultrastructure using cryo preparation in this integrated system.

Main Methods:

  • Design and construction of a complete cryo transfer flange for cryogenic specimen control.
  • Integration of the cryo transfer system with the EMRAM instrument (Raman spectrometer and XL-30 ESEM).
  • Acquisition of Raman spectra for polystyrene beads and sucrose at ambient and cryogenic temperatures, and during a heating ramp.

Main Results:

  • Cryogenic preparation preserved the chemical composition and ultrastructure of model specimens.
  • Raman spectra showed minimal differences in main peak positions between ambient and cryogenic conditions.
  • A significant sharpening of spectral bands in the 800-400 cm(-1) region was observed at cryogenic temperatures due to reduced intermolecular interactions.
  • Enhanced visibility of lower frequency modes was noted at cryogenic temperatures.

Conclusions:

  • The developed cryo transfer system effectively enables cryogenic control for combined Raman microspectrometry and SEM.
  • Cryogenic conditions enhance Raman spectral resolution by reducing intermolecular interactions, facilitating more detailed analysis.
  • This integrated approach significantly advances the noninvasive compositional analysis of delicate samples in electron microscopy.