Hello,
I would like to ask whose are studying about sonication of the alpha-synuclein preformed fibrils. The preformed fibrils are commercially available but in order to use for disease models, it must be prepared in a suitable particle size, which is less than 50 nanometers. I was searching publications but it's quite rare that a researcher describes the exact procedure and, of course, there are also differences between researchers and equipments which prevents standardization.
What is the pulse, duration, interval and other settings for sonication of the alpha-synuclein preformed fibrils into less than 50 nm particles, with probe sonicator?
Dear Halit Guner Orhan,
Here are some parameters you can try:
1. Sonicate for 30s (0.5s pulse on/off) at 10% amplitude using a Branson Digital sonifier
2. Sonicate using an ultrasonicating homogenizer (300VT; Biologics, Inc., Manassas, VA) with the pulser set at 20% and power output at 30% for 60 pulses at 1s each
3. Sonicate using either QSonica XL-2000 at power level 2 for a total of 30 pulses (1s each), or Fisher Scientific 120 Sonic Dismembrator equipped with CL-18 microtip (20% power)
References:
1. Kim, S., Kwon, S. H., Kam, T. I., Panicker, N., Karuppagounder, S. S., Lee, S., Lee, J. H., Kim, W. R., Kook, M., Foss, C. A., Shen, C., Lee, H., Kulkarni, S., Pasricha, P. J., Lee, G., Pomper, M. G., Dawson, V. L., Dawson, T. M., & Ko, H. S. (2019). Transneuronal Propagation of Pathologic α-Synuclein from the Gut to the Brain Models Parkinson's Disease. Neuron, 103(4), 627–641.e7.
2. Duffy, M. F., Collier, T. J., Patterson, J. R., Kemp, C. J., Luk, K. C., Tansey, M. G., Paumier, K. L., Kanaan, N. M., Fischer, D. L., Polinski, N. K., Barth, O. L., Howe, J. W., Vaikath, N. N., Majbour, N. K., El-Agnaf, O., & Sortwell, C. E. (2018). Lewy body-like alpha-synuclein inclusions trigger reactive microgliosis prior to nigral degeneration. Journal of neuroinflammation, 15(1), 129.
3. Fan, R. Z., Guo, M., Luo, S., Cui, M., & Tieu, K. (2019). Exosome release and neuropathology induced by α-synuclein: new insights into protective mechanisms of Drp1 inhibition. Acta neuropathologica communications, 7(1), 184.
Hope this helps!
Set up can differ if you are using a water bath or tip sonicator. We use Branson with 30 % output, as described at Article Organotypic slice culture model demonstrates inter-neuronal ...
you may also would like to check this paper if you haven't
doi: 10.3233/JPD-171248
Sarivin Vanan Sara Elfarrash Thank you for all replies. I will consider your suggestions. Also, I would like to add another Q&A about similar issue.
https://www.researchgate.net/post/About_alpha-synuclein_preformed_fibril_sonication