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Flow Cytometry01:23

Flow Cytometry

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The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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Related Experiment Video

Updated: Jun 12, 2025

Asymmetrical Flow Field-Flow Fractionation for Sizing of Gold Nanoparticles in Suspension
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Asymmetrical Flow Field-Flow Fractionation for Sizing of Gold Nanoparticles in Suspension

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Going hi-res in bulk: flowless multiangle dynamic light scattering for detection on asymmetric flow field flow

Lavinia Rita Doveri1, Giorgia Dal Pan1, Giovanni Tomaselli1

  • 1inLAB, Department of Chemistry, University of Pavia, Via Taramelli 12, Pavia, 27100, Italy. ydf@unipv.it.

Journal of Materials Chemistry. B
|May 23, 2025
PubMed
Summary

A new method combining asymmetric flow field flow fractionation (AF4) and multiangle dynamic light scattering (MADLS) accurately measures nanoparticle size without calibration. This technique offers a universal approach for nanomedicine quality control.

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Area of Science:

  • Nanomedicine
  • Analytical Chemistry
  • Materials Science

Background:

  • Nanomedicine requires precise analytical methods for regulatory compliance and safety.
  • Accurate particle size and distribution measurements are crucial for nanomedicine R&D and quality control.
  • Existing methods may require calibration materials, limiting their applicability and affordability.

Purpose of the Study:

  • To develop and validate a novel analytical approach for nanoparticle size determination.
  • To combine multidetector asymmetric flow field flow fractionation (MD-AF4) with multiangle dynamic light scattering (MADLS) for accurate sizing without calibration.
  • To evaluate the method's performance on various nanoparticles relevant to nanomedicine, environmental, and drug delivery applications.

Main Methods:

  • Utilized a combined MD-AF4 and MADLS system for nanoparticle characterization.
  • Employed model liposome formulations, prepared via lipid thin-film extrusion, for benchmarking.
  • Validated results against cryogenic transmission electron microscopy (cryoTEM) and standalone MADLS.
  • Investigated the influence of sample concentration on analytical bias in AF4-DLS measurements.
  • Extended analysis to polystyrene and polymethyl methacrylate particles.

Main Results:

  • Achieved accurate particle size determination for bioinspired colloids and polymeric particles without calibration standards.
  • Demonstrated that analytical bias in in-flow DLS measurements follows a universal law, independent of nanoparticle material.
  • The combined AF4-MADLS approach, with fraction collection, provided high-resolution size distributions and accurate number concentration.
  • Liposomes exhibited high monodispersity and long-term stability, confirmed by cryoTEM and MADLS.

Conclusions:

  • The proposed MD-AF4-MADLS method offers a robust, calibration-free solution for nanoparticle sizing.
  • The findings suggest a universal behavior in DLS-based sizing bias, simplifying analytical method development.
  • This approach enhances analytical tools for nanomedicine, potentially leading to wider adoption of AF4-DLS instrumentation.