featuring the Echo Acoustic Technology
TITLES and AUTHORS
Epigenetics continues to emerge as an important target class for drug discovery and cancer research. As programs scale to evaluate many new targets related to epigenetic expression, new tools and techniques are required to enable efficient and reproducible high-throughput epigenetic screening. The combination of robust and precise instrumentation from Labcyte and BMG LABTECH enable the miniaturization and automation of PerkinElmer® AlphaLISA epigenetic assay kits without compromising performance and data quality.
In this proof of concept study (POC), the Ovation Single Cell RNA-Seq System miniaturization was validated by examining libraries prepared and sequenced at traditional and miniaturized reaction volumes. Additionally, the sample and reagent transfers were automated using the Echo® acoustic liquid handling technology. Echo liquid handlers transfer a wide range of fluids without contact of tips or recalibration between fluid types. The industry leading accuracy and precision of Echo liquid handlers at microliter and nanoliter volumes in combination with the NuGEN Ovation Single Cell RNA-Seq System increases library preparation throughput while reducing the costs. This enables a broader application of transcriptome analysis with NGS.
The Labcyte® MicroClime environmental lid minimizes edge effects and preserves the concentration of solutions in microplate wells. Unlike standard microplate lids, the MicroClime lid incorporates a unique, fluid-absorbing matrix. The absorbed fluid creates a vapor barrier that protects the sample from the exterior environment, especially near the edges of the microplate. The MicroClime lid is easy to use—simply place it on or lift it off a microplate. No specialized tools are required, and it is compatible with robotic plate lidding systems. The lid can be filled with many different fluids, using either a hand-held syringe or automated bulk fillers.
When filled with anhydrous DMSO, the MicroClime lid prevents evaporation and hydration of DMSO in compound library plates during storage. It also stops evaporation of ultra-low-volume DMSO droplets in screening assays. The lid can also be filled with water to prevent evaporation in aqueous assays.
Labcyte Echo liquid handlers transfer compounds and reagents at nanoliter volumes without the use of tips or contact. This capability provides the precision and accuracy to miniaturize biochemical and cell-based assays for screening in 1536-well microplates. AlphaLISA technology is a faster, no-wash alternative to traditional ELISA assays for biomarker and cytokine detection. With high sensitivity and low background, these assays can be readily adapted to miniaturization with the Echo liquid handler. To complement the performance of the Echo liquid handler, the BMG PHERAstar® FS plate reader provides excellent sensitivity and throughput for miniaturized, high density assays. In this application note, we use a TNF-α AlphaLISA assay to validate the performance of a miniaturized assay.
Acoustic liquid handling utilizes high-frequency sound waves that are focused on the surface of a fluid to eject nano- and picoliter volumes with high accuracy and precision (Figure 1). The Echo® 550 and Echo® 555 liquid handlers transfer 2.5 nL droplets from 384-well and 1536-well microtiter plates, across a wide range of fluids up to 1.25 μL/s.
The new Echo® 525 liquid handler increases the transfer droplet volume 10-fold to 25 nL, which allows for transfer rates of 5 μL/s and enables workflows for life science applications that were previously less practical at slower rates. To enable the transfer of large quantities of fluid at this faster transfer rate we are developing a companion consumable, the multi-well Reagent Reservoir (RES), for use in these various applications.
Recently identified associations between variants of cancer genes and drug resistance have increased the value for comprehensive drug sensitivity screening in combination with molecular profiling of cancer cells. This in turn, has led to a demand for improvements to screening throughput and efficiency. Echo® Liquid Handlers use acoustic energy to provide high throughput, non-contact, liquid handling for a range of applications. Non-contact transfer avoids the risk of cross-contamination, eliminates tip costs, and facilitates the progression to high density assay formats. Echo Liquid handlers precisely and accurately transfer 2.5 nL droplets of sample and reagent, which enables the miniaturization of biochemical screens. This poster discusses the implementation of miniaturized drug sensitivity screening, at the Institute of Molecular Medicine in Finland (FIMM), with assay-ready plates produced by the Echo liquid handler.
Liquid handling and dilution processes profoundly influence estimates of biological activity. We compared tip-based serial dilution and dispensing versus acoustic dispensing with direct dilution. Published data show results differ by orders of magnitude with no correlation or ranking of datasets. Pharmacophores generated from data derived by both acoustic and tip-based transfer differ significantly. The acoustic dispensing-derived pharmacophore correctly identified active compounds in a subsequent test set where the tip-based method failed completely. Acoustic data generates a pharmacophore containing two hydrophobic features, one hydrogen bond donor and one hydrogen bond acceptor and is consistent with X-ray crystallography studies. In contrast, the tip-based data suggest a pharmacophore with two hydrogen bond acceptors, one hydrogen bond donor and no hydrophobic features. This pharmacophore is inconsistent with the X-ray crystallographic studies. In short, tip-based dispensing processes are an important source of error in high-throughput screening. These findings have far-reaching implications in biological research.
Small molecule screening against kinases is commonly used to discover drug candidates that target various human diseases. The sheer number of small molecules tested and new targets identified every year drives screening programs to push the limits of throughput and efficiency. By combining the nanoliter-scale and non-contact Labcyte Echo 525 liquid handler with the robust and ultra-sensitive Promega ADP-Glo assay, high throughput screening can be improved. This poster demonstrates the reproducibility of inhibition and IC50 determinations from a high throughput screen performed with the Echo 525 liquid handler. In this miniaturized and high throughput format high quality and reliable data can be achieved with reduced sample and reagent volumes.
PerkinElmer AlphaLISA® technology is a faster, no-wash alternative to traditional ELISA assays for biomarker and cytokine detection. With high sensitivity and low background, these assays can be miniaturized to low volumes and adapted to automation. Labcyte Echo liquid handlers transfer compounds and reagents at nanoliter volumes without the use of tips or contact. This capability provides precision and accuracy to miniaturize biochemical assays for screening in 1536-well microplates. To complement the performance of Echo liquid handlers, the BMG PHERAstar FS plate reader provides excellent sensitivity and throughput for miniaturized, high density assays.
The advent of a new technology often brings far-reaching benefits well beyond the initial perception of its key advantages. Acoustic dispensing, for example, with the ability to have highly controlled droplet transfers at 2.5nL increments with exquisite precision and accuracy, does much more than save money by eliminating pipets. It enables new work flows, including those in which multiple pharmaceutical firms have seen dramatic differences in the true success or failure of compounds. This article shows how the use of acoustic dispensing and direct dilution leads to the creation of better compound structures and the potential for faster development of new drugs.
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