3D cell culture-based drug discovery
Conclusion: Automated liquid handling enhances both the throughput and reproducibility of drug screening workflows using 3D cell cultures.
Magnetic three-dimensional cell culture (M3D) offers a way to address some of the current challenges in 3D cell culture, allowing quick and consistent generation of spheroids. This approach can be automated using the Tecan Fluent liquid handler to produce uniformly sized spheroids in flat-bottomed microplates suitable for downstream high throughput screening applications. In this application, HeLa cells were seeded and incubated using Fluent. To illustrate the efficacy of the resulting spheroids in compound screening, a D300e Digital Dispenser was used to directly dispense drug stock solutions into wells containing the spheroids to generate the dose-response curve. Brightfield imaging and luminescence measurements were performed with the Tecan Spark Cyto 600 multimode reader to conduct imaging-based quality control throughout the growth phase.
The combination of the Fluent, D300e dispenser and M3D significantly accelerated the workflow, reducing time and material consumption. Imaging-based evaluation using the Spark Cyto allowed growth monitoring and viability assessment to be performed with a single device, improving data acquisition efficiency through multiplexing. The use of a 384-well microplate expanded throughput, allowing simultaneous analysis of a larger number of samples while reducing time and material consumption. Combining controlled spheroid formation with automated liquid handling and high-quality imaging enhances both the throughput and reproducibility of drug screening workflows using 3D cell cultures, with applications in drug discovery and biomedical research.
Entire application note: Automated spheroid production by magnetic cell assembly for screening applications
Automated DNA extraction
Conclusion: Automated DNA extraction workflow using liquid handling provides quickly enough high-quality DNA for e.g. NGS sequencing or qPCR analysis.
DNA extraction workflow for Ortho-Analytic’s Calex NGS stool sampling tubes was automated using the Tecan DreamPrep NAP liquid handler with the ZymoBIOMICS 96 MagBead DNA Kit and the results compared with the ZymoBIOMICS DNA miniprep manual extraction kit. After elution, onboard DNA quantification was performed without any sample loss using UV absorbance-based measurements on the integrated Frida Reader. This allowed normalization of all samples for further downstream applications as required. A standard fluorometer was used to compare DNA quantification results. The extracted DNA was sequenced for comparison of manual and automated extraction procedures.
The results showed that the automated DNA extraction workflow, using ZymoBIOMICS 96 MagBead DNA kit, delivers sufficient amounts of DNA for further downstream applications, with a significant reduction in production time and higher sample throughput. The integrated Frida Reader operated close to its detection limit, but still delivered concentration measurements that correlated with the highly specific and sensitive fluorometric measurements. The use of Frida Reader allowed DNA concentration measurements to be performed directly on the instrument, without wasting precious material. The subsequent normalization step saved further hands-on time, increasing daily throughput. Side-by-side comparison of manual and automated DNA extraction revealed excellent correlation between sequencing results for both methods.
Entire application note: Simplified sample collection and automated DNA extraction for high throughput sequencing and intestinal gut microbiome analysis
Automated sample preparation for plasma proteomics
Conclusion: Automated liquid handling with integrated positive pressure module speeds up the proteomics sample preparation process.
Plasma proteome profiling workflow for PreOmics’ ENRICH-iST was fully automated using Tecan Fluent liquid handler with integrated Resolvex i300 positive pressure module. This allowed direct peptide clean-up and drying on the Fluent system. The streamlined process started with plasma samples and yielded dried peptides. The automated workflow was evaluated through a multi-day analysis to assess its reproducibility and robustness, and further tested in a study examining age-related plasma proteome differences between two age groups.
The integration of the on-deck Resolvex i300 positive pressure module enabled complete end-to-end automation of the sample preparation process, speeding up plasma sample preparation and minimizing manual steps. The total time from plasma to dried peptides is spanning 6 hours, enabling the preparation of 96 samples within a single working day.
Entire application note: Simplified, automated sample preparation for plasma proteomics