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Related Concept Videos

Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been developed.
Overview of Electron Microscopy01:25

Overview of Electron Microscopy

The wavelengths of visible light ultimately limit the maximum theoretical resolution of images created by light microscopes. Most light microscopes can only magnify 1000X, and a few can magnify up to 1500X. Electrons, like electromagnetic radiation, can behave like waves, but with wavelengths of 0.005 nm, they produce significantly greater resolution up to 0.05 nm as compared to 500 nm for visible light. An electron microscope (EM) can create a sharp image that is magnified up to 2,000,000X.
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
Accelerated...
Spectrophotometry: Introduction01:16

Spectrophotometry: Introduction

Spectrophotometry is the quantitative measurement of the absorption, reflection, diffraction, or transmission of electromagnetic radiation through a material as a function of the intensity and wavelength of the radiation. A spectrophotometer is a device used to measure the change in the radiation intensity caused by its interaction with the material.
The essential components of a spectrophotometer include a source of electromagnetic radiation, a slot for placing a material to be analyzed, and a...

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Related Experiment Video

Updated: May 27, 2026

Measurement of Scattering Nonlinearities from a Single Plasmonic Nanoparticle
15:06

Measurement of Scattering Nonlinearities from a Single Plasmonic Nanoparticle

Published on: January 3, 2016

Evaluation of a spectrally resolved scattering microscope.

Michael Schmitz1, Thomas Rothe, Alwin Kienle

  • 1Institut für Lasertechnologien in der Medizin und Meßtechnik, 89081 Ulm, Germany.

Biomedical Optics Express
|November 18, 2011
PubMed
Summary

A new scattering microscope accurately measures single cell and biological microstructure sizes using light scattering. This method, validated against traditional techniques, offers precise diameter determination with minimal error for advanced biological research.

Keywords:
(180.0180) Microscopy(290.1350) Back scattering(290.2200) Extinction(290.4020) Mie theory(290.5850) Scattering, particles(300.6550) Spectroscopy, visible

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Area of Science:

  • Biophysics
  • Optical Microscopy
  • Nanotechnology

Background:

  • Accurate size measurement of microstructures is crucial for biological research.
  • Traditional methods may have limitations in resolution or sample manipulation.
  • Developing novel microscopy techniques is essential for advancing cell biology.

Purpose of the Study:

  • To develop and validate a spectrally resolved scattering microscope for investigating single cells and biological microstructures.
  • To compare the performance of the new scattering microscope with a conventional collimated transmission setup.
  • To assess the accuracy and precision of light scattering measurements for determining particle size.

Main Methods:

  • Development of a spectrally resolved scattering microscope setup.
  • Validation of the setup using homogenous polystyrene spheres.
  • Application of Mie theory for accurate diameter determination.
  • Comparative analysis with a collimated transmission setup for size distribution analysis.

Main Results:

  • The scattering microscope successfully determined the diameters of 150 single polystyrene spheres.
  • Both the scattering microscope and the collimated transmission setup yielded mean diameters and standard deviations with a statistical error of less than 1nm.
  • Systematic errors between the two methods were found to be in agreement within the measurement accuracy.

Conclusions:

  • The developed spectrally resolved scattering microscope is a viable and accurate tool for measuring the size of microstructures.
  • Light scattering measurements, particularly when spectrally resolved, provide precise and reliable size information comparable to established methods.
  • This technology has potential applications in cell biology, nanotechnology, and materials science where precise size characterization is critical.