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    A new variable gain calibration for Laser Transmission Spectroscopy (LTS) significantly improves nanoparticle size measurement accuracy. This method enhances precision for nanoparticle characterization across various scientific and industrial fields.

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    Area of Science:

    • Nanotechnology and Materials Science
    • Analytical Chemistry
    • Biotechnology

    Background:

    • Accurate nanoparticle size and concentration are crucial for materials science, biotechnology, pharmaceutics, and food industries.
    • Laser Transmission Spectroscopy (LTS) is a unique single-measurement technique for determining nanoparticle size and concentration.

    Purpose of the Study:

    • To introduce a novel variable gain calibration procedure for LTS instruments.
    • To enhance the precision of nanoparticle size determination using LTS.
    • To validate the new calibration method with standard nanoparticles and biological samples.

    Main Methods:

    • Development of a variable gain calibration procedure for LTS.
    • Utilization of a custom-designed tunable-gain, dual-channel, dual-phase Lock-In Amplifier (LIA).
    • Measurement of laser beam intensities through nanoparticle suspensions and a reference path using Si photodiodes.

    Main Results:

    • The new LTS calibration procedure reduces experimental uncertainty in particle size by an order of magnitude compared to the double ratio technique.
    • Validation with NIST standard polystyrene nanoparticles demonstrated sustained accuracy.
    • Characterization of extracellular vesicles from orange juice (25 nm) and their aggregates (340 nm) was achieved in a single LTS measurement.

    Conclusions:

    • The variable gain calibration significantly improves the accuracy and reliability of LTS for nanoparticle characterization.
    • This advanced LTS method is effective for analyzing biological nanostructures, including extracellular vesicles.
    • The enhanced LTS technique offers a powerful tool for research and industrial applications requiring precise nanoparticle analysis.