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Related Concept Videos

Cryo-electron Microscopy01:28

Cryo-electron Microscopy

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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...
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Updated: Jun 5, 2025

Cryo-Electron Microscopic Grid Preparation for Time-Resolved Studies using a Novel Robotic System, Spotiton
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Passage through micro-sprayer increases functional activity - implications for activity assays in time-resolved

Priyanka Garg, Xiangsong Feng, Swastik De

    Biorxiv : the Preprint Server for Biology
    |December 16, 2024
    PubMed
    Summary

    This study found that ribosome functional competence assays using micro-sprayers show an unexpected increase in activity, not a decrease. Mechanical agitation, like sonication, appears to be the key factor influencing ribosome behavior.

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    Area of Science:

    • Biochemistry
    • Molecular Biology
    • Structural Biology

    Background:

    • Ribosomes are essential for protein synthesis.
    • Assays are used to measure ribosome functional competence.
    • Microfluidic devices are increasingly used in biological studies.

    Purpose of the Study:

    • To validate an assay measuring ribosome functional competence after passing through a micro-sprayer.
    • To investigate the effect of micro-spraying on ribosome activity.

    Main Methods:

    • Monitoring Green Fluorescent Protein (GFP) production via fluorescence.
    • Utilizing a micro-sprayer assay to assess ribosome function.
    • Applying sonication as a method of mechanical agitation.

    Main Results:

    • An unexpected increase in GFP production was observed, contrary to the expected decrease.
    • Ribosome heterogeneity or partial aggregation in solution may explain the findings.
    • Sonication led to a significant increase in ribosome functional activity.

    Conclusions:

    • The micro-sprayer assay's validity is questionable due to observed increases in ribosome activity.
    • Mechanical agitation is a critical factor influencing ribosome function in these assays.
    • Findings impact the design and interpretation of time-resolved cryo-electron microscopy (cryo-EM) experiments using microfluidic chips.