Integration of segmented microflow chemistry and online HPLC/MS analysis on a microfluidic chip system enabling enantioselective analyses at the nanoliter scale

Piendl, Sebastian K.; Schönfelder, Thomas; Polack, Matthias; Weigelt, Laura; Zwaag, T. van der; Teutenberg, Thorsten; Beckert, Erik; Belder, Detlev

doi:10.1039/d1lc00078k

Cited by 27 publications

(23 citation statements)

References 64 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…[14] For microfluidic operations, e.g., mixing or cell encapsulation, we applied monolithic fused-silica glass chips that were microstructured via selective laser-induced etching. [15] This enabled a dead volume-free connection of standard capillaries with an outer diameter (o.d.) of 360 μm and allowed the generation of uniform droplets of constant volumes in the nL range.…”

Section: Resultsmentioning

confidence: 99%

Quantification of Biocatalytic Transformations by Single Microbial Cells Enabled by Tailored Integration of Droplet Microfluidics and Mass Spectrometry

et al. 2022

Self Cite

View full text Add to dashboard Cite

Improving the performance of chemical transformations catalysed by microbial biocatalysts requires a deep understanding of cellular processes. While the cellular heterogeneity of cellular characteristics, such as the concentration of high abundant cellular content, is well studied, little is known about the reactivity of individual cells and its impact on the chemical identity, quantity, and purity of excreted products. Biocatalytic transformations were monitored chemically specific and quantifiable at the single-cell level by integrating droplet microfluidics, cell imaging, and mass spectrometry. Product formation rates for individual Saccharomyces cerevisiae cells were obtained by i) incubating nanolitresized droplets for product accumulation in microfluidic devices, ii) an imaging setup to determine the number of cells in the droplets, and iii) electrospray ionisation mass spectrometry for reading the chemical contents of individual droplets. These findings now enable the study of whole-cell biocatalysis at single-cell resolution.

show abstract

Section: Resultsmentioning

confidence: 99%

Quantification of Biocatalytic Transformations by Single Microbial Cells Enabled by Tailored Integration of Droplet Microfluidics and Mass Spectrometry

et al. 2022

Self Cite

View full text Add to dashboard Cite

show abstract

“…Für mikrofluidische Operationen, z.B. Mischen oder Einkapseln von Zellen, verwendeten wir monolithische Glas‐Chips aus Quarzglas, die durch selektives laserinduziertes Ätzen mikrostrukturiert wurden [15] . Dies ermöglichte eine totvolumenfreie Anbindung von Standardkapillaren mit einem Außendurchmesser (AD) von 360 μm, wodurch monodisperse Tröpfchen mit konstanten Volumina im nL‐Bereich erzeugt werden konnten.…”

Section: Ergebnisse Und Diskussionunclassified

Quantifizierung biokatalytischer Umwandlungen durch einzelne mikrobielle Zellen mittels maßgeschneiderter Integration von Tröpfchenmikrofluidik und Massenspektrometrie

et al. 2022

Self Cite

View full text Add to dashboard Cite

Die Verbesserung der Effizienz von chemischen Umwandlungen, die durch mikrobielle Biokatalysatoren katalysiert werden, erfordert ein tiefes Verständnis von zellulären Prozessen. Während die Heterogenität zellulärer Merkmale, wie die Konzentration des reichlich vorhandenen Zellinhalts, gut untersucht ist, ist wenig über die Reaktivität einzelner Zellen und ihre Auswirkung auf die chemische Identität, Menge und Reinheit der sekretierten Produkte bekannt. Biokatalytische Umwandlungen wurden chemisch spezifisch und quantifizierbar auf der Einzelzellebene durch die Integration von Tröpfchenmikrofluidik, Zellbildgebung und Massenspektrometrie überwacht. Die Produktbildungsraten für einzelne Saccharomyces cerevisiae-Zellen wurden durch i) Inkubation von nanolitergroßen Tröpfchen zur Produktakkumulation in Mikrofluidik-Plattformen, ii) eine Bildgebungseinheit zur Bestimmung der Zellanzahl in den Tröpfchen und iii) Elektrospray-Ionisations-Massenspektrometrie zur Messung des chemischen Inhalts einzelner Tröpfchen ermittelt. Diese Erkenntnisse ermöglichen nun die Untersuchung der Biokatalyse ganzer Zellen bei einer Einzelzellauflösung.

show abstract

“…Following the splitting of the large volumes of dialysate into picoliter to nanoliter volume droplets, analytical methods were adapted to analyze their content and a library of highly-sensitive solutions, unperturbed by surfactants and oil 27 , was developed. This benefitted from the rich literature of droplet microfluidics (e.g., droplet generation 28 , picoinjection 29 , phase extraction [30][31][32] , etc.). These solutions, which were extensively covered in recent reviews [33][34][35][36] , comprise in-flow electrochemical chips 37,38 , fluorescent assays 23,39,40 and include many developments of electrophoretic 30,41 and mass spectrometry methods 26,[42][43][44][45][46][47][48] .…”

Section: Introductionmentioning

confidence: 99%

On demand nanoliter sampling probe for collection of brain fluid

Teixidor

Novello

Ortiz

et al. 2022

Preprint

View full text Add to dashboard Cite

Continuous fluidic sampling systems allow collection of brain biomarkers in vivo. Here, we propose a new sampling paradigm, Droplet on Demand (DoD), implemented in a microfabricated neural probe. It allows sampling droplets loaded with molecules from the brain extracellular fluid punctually, without the long transient equilibration periods typical of continuous methods. It uses an accurate fluidic sequence and correct operation is verified by the embedded electrodes. As a proof of concept, we demonstrated the application of this novel approach in vitro and in vivo, to collect glucose in the brain of mice, with a temporal resolution of 1-2 minutes and without transient regime. Absolute quantification of the glucose level in the samples was performed by direct infusion nanoelectrospray ionization Fourier transform mass spectrometry (nanoESI-FTMS). By adjusting the diffusion time and the perfusion volume of DoD, the fraction of molecules recovered in the samples can be tuned to mirror the tissue concentration at accurate points in time. This makes quantification of biomarkers in the brain possible within acute experiments of only 20 to 120 minutes. DoD provides a complementary tool to continuous microdialysis and push-pull sampling probes. The advances allowed by DoD will benefit quantitative molecular studies in the brain, namely for molecules involved in volume transmission or for protein aggregates that form in neurodegenerative diseases over long periods.

show abstract

Integration of segmented microflow chemistry and online HPLC/MS analysis on a microfluidic chip system enabling enantioselective analyses at the nanoliter scale

Abstract: In this work, we introduce an approach to merge droplet microfluidics with a HPLC/MS functionality on a single chip to analyze the contents of individual droplets. This is achieved by...

Cited by 27 publications

References 64 publications

Quantification of Biocatalytic Transformations by Single Microbial Cells Enabled by Tailored Integration of Droplet Microfluidics and Mass Spectrometry

Quantification of Biocatalytic Transformations by Single Microbial Cells Enabled by Tailored Integration of Droplet Microfluidics and Mass Spectrometry

Quantifizierung biokatalytischer Umwandlungen durch einzelne mikrobielle Zellen mittels maßgeschneiderter Integration von Tröpfchenmikrofluidik und Massenspektrometrie

On demand nanoliter sampling probe for collection of brain fluid

Contact Info

Product

Resources

About