Alpha-Synuclein Fragmentation using the VialTweeter Sonicator

α-synuclein fibrils and ribbons are fragmented in scientific research to generate smaller fibril snippets or even individual protein molecules, which can be more readily analyzed using various experimental techniques. The VialTweeter Sonicator is one of the most commonly used ultrasonicators for efficient and reliable alpha-synuclein fragmentation.

α-Synuclein in Research

Alpha-synuclein fibrils are protein aggregates that are strongly associated with neurodegenerative disorders such as Parkinson disease and certain forms of dementia, including dementia with Lewy bodies. Research focused on alpha-synuclein fibrils aims to understand their role in disease progression and develop potential therapeutic interventions. By breaking down alpha-synuclein fibrils into smaller fragments, researchers can investigate their specific structural features. For instance, fragmenting alpha-synuclein fibrils enables researchers to investigate their interactions with other molecules, such as proteins, lipids, or small molecules. By producing smaller fragments, the binding sites and affinity for these interacting partners can be probed more effectively. Smaller α-Syn fribrils and ribbons may also show altered toxicity and biochemical effects. Therefore, a reliable and efficient fragmentation technique, which produces reproducible results in a quick and simple sample treatment, is crucial.
Ultrasonic Alpha-Syn Fragmentation: The VialTweeter sonicator is the established ultrasonic sample prep system that sonicates up to 10 vials simultaneously under exactly the same conditions. Programmable settings allow for the simple and quick re-run of the same experiments, which gives highly reliable and reproducible results in alpha-synuclein fibril fragmentation.

α-Synuclein Sample Preparation with a Sonicator

One approach to studying alpha-synuclein fibrils involves their extraction and fragmentation using techniques such as sonication. Sonication is a process that uses high-intensity, low-frequency ultrasound waves to disrupt and break down protein aggregates, leading to the release of smaller fibrils or individual protein molecules. The sonicator VialTweeter is a commonly used device in α-synuclein-related research studies for this purpose.

Numerous research studies describe precise sample preparation protocols of alpha-synuclein fibrils sonication, which use the Hielscher VialTweeter for efficient and reliable α-synuclein fibril fragmentation. By fragmenting the fibrils ultrasonically, researchers can analyze the resulting products and examine their structure, toxicity, and interactions with other molecules. This research provides important insights into the mechanisms underlying neurodegeneration and potentially identify new therapeutic targets. The well established α-synuclein sonication protocols using the VialTweeter sonicator allow for reliable and reproducible results.

Upper images: unfragmented alpha-synuclein fibrils
Lower images: Ultrasonically fragmented alpha-synuclein fibrils with the VialTweeter sonicator
(study and images: ©Dieriks et al., 2022)

Ultrasonic Fragmentation of α-Synuclein Fibrils – Protocols

Since numerous researchers use the VialTweeter sonicator as preferred fragmentation technique to produce uniform α-synuclein fibril fragments, established protocols are readily available. Below you can find a few exemplary fragmentation protocols.
 

 
ClearTau seed preparation: The ClearTau fibrils were diluted to 10 μM in dH2O and sonicated at 70 % amplitude for 50 s with 1 s ON 1 s OFF cycle in-tube using the sonicator UP200St with VialTweeter. Seeds were characterized by electron microscopy.
ThS fluorescence measurement: The ClearTau fibrils were diluted to 2.5 μM in dH2O and sonicated at 70 % amplitude for 50 s with 1 s ON 1 s OFF cycle in-tube using UP200St with VialTweeter. 2.5 μM full-length Tau 4R2N monomer was used as a control. To the 100 μl reaction, 100 μl of ThS (10 μM) were added, yielding final protein concentrations of 1.25 μM. Single-time point ThS fluorescence was measured using 96 well clear bottom plates set up in FLUOstar Omega microplate reader with excitation at 445 nm and emission at 485 nm was recorded.
(cf. Limorenko et al., 2023)
 
Uniform α-synuclein length using sonication: The length heterogeneity of α-syn fibrils and ribbons was reduced by sonication for 20 min on ice in 2-ml Eppendorf tubes in a VialTweeter set at 75% amplitude, 0.5 s pulses.
(cf. Bousset et al., 2013)
 
The quality control of human recombinant monomeric WT or S129A a-Syn and the fibrillar polymorphs they generate as well as that of a-Syn 1–110 was carried out. Subsequently, the fibrillar polymorphs were fragmented by sonication for 20 min in 2-mL Eppendorf tubes in the VialTweeter ultrasonicator to generate fibrillar particles with an average size of 42–52 nm that are suitable for endocytosis.
(cf. Shrivastava et al., 2020)
 

Characterization of five fibrillar α-Syn polymorphs. (A) Transmission electron micrographs of negatively stained α-Syn fibrillar polymorphs fibrils, ribbons, fibrils-91, fibrils-65, and fibrils-110 before (upper lane) and after fragmentation with the VialTweeter (lower lane) are shown. (B) Length distribution of the fragmented fibrillar polymorphs is shown. The number (n) of fibrillar assemblies the histograms were derived from is indicated.
(study and images: Shrivastava et al., 2020)

 
The α-Syn fibrils were fragmented by sonication for 20 min in 2 ml Eppendorf tubes in a VialTweeter to generate fibrillar particles with an average size 42–52 nm as assessed by TEM analysis.
(cf. Negrini et al., 2022)
 
The resuspended fibrils91 (in PBS) were fragmented prior to addition to cell cultures by sonication for 20 min in 2 mL Eppendorf tubes using the Vial Tweeter sonicator, aliquoted, flash frozen in liquid nitrogen and stored until use at -80 ̊C.
(cf. Vajhøj et al., 2021)
 

VialTweeter and Lab Sonicators for α-Syn Fragmentation

Hielscher Ultrasonics is worldwide recognized as leading manufacturer of cutting-edge ultrasonicators. Trusted and established in major research labs worldwide, our ultrasonicators offer unparalleled quality and performance for your critical experiments.
With the Hielscher VialTweeter and any other Hielscher sonicator, you will experience unparalleled user comfort, as they are thoughtfully engineered for reproducible results, ease of use and seamless operation. With automatic data recording features, you can focus on your research while our ultrasonicators meticulously record crucial data for reproducibility and precision.
Achieve consistent and reproducible results with confidence!
Our ultrasonicators are engineered and manufactured in Germany. German precision and highest engineering quality to ensure reliable and accurate sample preparation such alpha-synuclein fibril fragmentation every time. No more worries about inconsistent outcomes – our ultrasound technology ensures your research remains at the forefront of scientific advancement.
But that is not all! Our commitment to excellence extends beyond our products. We take pride in our exceptional customer service, providing expert support to address any queries or concerns you may have. Our dedicated team is here to assist you at every step of your research journey, ensuring you get the most out of our ultrasonicators.
Choose innovation, reliability, and exceptional user experience – choose our lab sonicators such as the VialTweeter for alpha-synuclein fibril fragmentation. Benefit from the already established protocols and join the ranks of leading researchers who trust our technology for their critical studies. Elevate your research and explore new frontiers in dementia and neurodegenerative disease research with our state-of-the-art ultrasonicators.

Design, Manufacturing and Consulting – Quality Made in Germany

Hielscher ultrasonicators are well-known for their highest quality and design standards. Robustness and easy operation allow the smooth integration of our ultrasonicators into industrial facilities. Rough conditions and demanding environments are easily handled by Hielscher ultrasonicators.

Hielscher Ultrasonics is an ISO certified company and put special emphasis on high-performance ultrasonicators featuring state-of-the-art technology and user-friendliness. Of course, Hielscher sonicators are CE compliant and meet the requirements of UL, CSA and RoHs.
 
The table below gives you an indication of the approximate processing capacity of our lab-size ultrasonicators: