Novel delivery and sample mixing for synchrotron diffraction experiments using acoustic levitation with multi-transducer arrays

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Dye, ER ORCID: https://orcid.org/0000-0001-5425-619X, 2022. Novel delivery and sample mixing for synchrotron diffraction experiments using acoustic levitation with multi-transducer arrays. PhD, Nottingham Trent University.

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Abstract

Acoustic levitation may utilise standing waves at ultrasonic frequencies to manipulate suspended substances and small objects in a contactless manner. These materials may be levitated in the positions in which the nodes are located, corresponding to positions of low acoustic pressure. In recent years, off the shelf transducer based acoustic levitators have been used for contactless manipulation of liquids. These systems benefit from requiring low power and low-cost components making acoustic levitation more accessible to the masses. Such a system was investigated in this work for presenting protein crystals, within their mother liquor, to the I24 beamline at Diamond Light Source for x-ray diffraction experiments. It was found that the crystals tended to sediment toward the bottom of the droplets, which were oblate in shape. The droplets which were levitated often became unstable and fell from their suspended position, or they would not detach from the pipette tip when they were being injected. To rectify this, a coating of silicone oil was added allowing the droplets to remain stable as well as limit the evaporation of the droplet whilst it was manually inserted and the area cleared of personnel before the x-ray beam was engaged. This silicone oil coating is non-crystalline and thus did not invalidate the results collected which showed the lysozyme crystal structure with a resolution of 1.69 A, confirming acoustic levitation as a good sample presentation method for these types of experiments. To remove the requirement for the silicone oil, a bespoke system was created named the DLS-Lev that allowed top-loading of the sample. The droplets of mother liquor which contained protein crystals were easily detached from the pipette tip into the traps within the DLS-Lev system owing to the increased strength of the traps in the modified design. This system, paired with an automated pipette, facilitated sample mixing experiments whilst the x-ray beam was engaged. The further development of the pipetting system was halted due to the COVID-19 pandemic. However, future work should see the permanent installation of these systems at the I24 beamline at Diamond Light Source, as well as additional bespoke acoustic levitators designed for the other beamlines specialising in the research of protein structure via x-ray scattering techniques.

Item Type:	Thesis
Description:	This research programme was carried out in collaboration with Diamond Light Source.
Creators:	Dye, E.R.
Date:	June 2022
Rights:	This work is the intellectual property of the author, Nottingham Trent University and Diamond Light Source Ltd. You may copy up to 5% of this work for private study, or personal, non-commercial research. Any re-use of the information contained within this document should be fully referenced, quoting the author, title, university, degree level and pagination. Queries or requests for any other use, or if a more substantial copy is required, should be directed in the owners of the Intellectual Property Rights.
Divisions:	Schools > School of Science and Technology
Record created by:	Linda Sullivan
Date Added:	07 Sep 2022 13:38
Last Modified:	07 Sep 2022 13:38
URI:	https://irep.ntu.ac.uk/id/eprint/46980