Droplet microfluidics for time-resolved serial crystallography.
IUCrJ
; 11(Pt 2): 237-248, 2024 Mar 01.
Article
em En
| MEDLINE
| ID: mdl-38446456
ABSTRACT
Serial crystallography requires large numbers of microcrystals and robust strategies to rapidly apply substrates to initiate reactions in time-resolved studies. Here, we report the use of droplet miniaturization for the controlled production of uniform crystals, providing an avenue for controlled substrate addition and synchronous reaction initiation. The approach was evaluated using two enzymatic systems, yielding 3â
µm crystals of lysozyme and 2â
µm crystals of Pdx1, an Arabidopsis enzyme involved in vitamin B6 biosynthesis. A seeding strategy was used to overcome the improbability of Pdx1 nucleation occurring with diminishing droplet volumes. Convection within droplets was exploited for rapid crystal mixing with ligands. Mixing times of <2â
ms were achieved. Droplet microfluidics for crystal size engineering and rapid micromixing can be utilized to advance time-resolved serial crystallography.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Arabidopsis
/
Microfluídica
Idioma:
En
Revista:
IUCrJ
Ano de publicação:
2024
Tipo de documento:
Article
País de afiliação:
Reino Unido