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Acoustic wave pipetting technology leader: ATS acoustic wave pipetting principle and application case
ATS was born in EDC, a low-profile US company focused on technology development, and was quickly adopted by companies such as GSK, Novartis, Merck, and Pfizer. The Broad Institute and the Berkeley National Key Laboratory also noted the use of ATS for quantitative PCR-based high-throughput drug screening and high-throughput mass spectrometry experiments.
What is the technology of ATS? Let us come to understand him.
Sound waves are almost everywhere, very common, but ATS can use it for liquid transfer. The ATS uses an acoustic probe to convert electrical energy into sound waves and listen to the reflection of sound waves to obtain information on the nature of the sample and the height of the liquid level. Based on the information obtained, the sonic probe automatically adjusts the position of the probe to precisely focus the energy to a point on the liquid surface to excite the cashier. The droplets fly to the upper container, and the droplets adhere to the surface of the container by surface tension or directly into the reaction system (see Figure 1). The ATS precisely controls the droplet size by adjusting the state of the energy, and arbitrarily adjusts the droplet volume between 1 nl and 50 nl according to the experimental needs. A larger volume of liquid transfer is achieved by the transfer of hundreds of droplets per second.
Figure 1: Schematic diagram of ATS acoustic wave pipetting
ATS can transfer any hole to any hole. During the pipetting process, the sample passes through the air at a rapid speed, without the need for tips, nozzles and tubing, without liquid residue, cross-contamination and blockage. The liquid to be transferred can be stored in 96, 384, 1536, and 3456 well plates, and the ATS can transfer liquid to any SBS/ANSI plate, even a glass slide, crystal plate, or chip. The accuracy and precision of ATS pipetting is much higher than traditional pipetting techniques, and the dead volume is less than 0.5 ul, which can maximize the use of valuable experimental samples.
At present, sonic pipetting technology has made achievements in the following fields: high-throughput drug screening, structural biology, molecular diagnosis, synthetic biology, precision medicine, high-throughput mass spectrometry. Now share a few application cases with you.
1. High-throughput drug screening : Innovating traditional experimental procedures, narrowing the reaction system, and achieving high-throughput, low-cost drug screening
In the IC50 experiment, the solubility of the mother liquor compound is high. In the conventional experimental procedure, the mother liquor is first transferred to a microplate and subjected to gradient dilution using a pipetting device. The diluted compound was then added to the experimental plate (Figure 2). It is difficult to avoid liquid residue in the tip of the entire pipetting process, resulting in deviations in experimental results.
Figure 2: Traditional gradient dilution pipetting process
The minimum droplet volume of the ATS is 1 nl. The direct dilution method (Figure 3) can be used to directly transfer the mother liquor of the compound to the experimental plate. This process can be assisted by ATS Cherry Pick and Dose-Response software. The accuracy and precision of ATS pipetting reduces the number of experimental iterations. A dead volume of 0.5 ul significantly reduces the volume of the solution required for the experiment, saving valuable samples and reagents.
Figure 3: ATS direct dilution method pipetting process
In the compound screening experiment, Bristol-Myers Squibb conducted a parallel comparison between sonic pipetting technology and traditional pipetting-based pipetting techniques. They surprisingly found that among the 1000 selected compounds, 108 of them were only using sonic waves. It is active when pipetting technology (Figure 4).
Figure 4: Comparison of screening results of Bristol-Myers Squibb's sonic pipetting and traditional pipetting methods
2. High-throughput quantitative PCR: automation, microsystems, very low CV
Quantitative PCR (qPCR) is widely used in gene expression analysis, genotyping, SNP analysis and other applications. At present, the commonly used reaction system for quantitative PCR is 10ul-20ul, and the quantitative PCR can be carried out in 1ul or even smaller reaction system by ATS sonic pipetting technology. ATS carries out nano-pumping of cDNA, Mix buffer, primers, and fluorescent reagents. The whole process requires no tips, no cross-contamination, and extremely low CV. Scientists at the Broad Institute of MIT and Harvard used ATS in conjunction with the Roche LightCycler® 1536 quantitative PCR machine to enable high-throughput drug screening based on quantitative PCR [1] . ATS can also be used in conjunction with a robotic arm to achieve 24-hour unattended high-throughput drug screening (Figure 5).
Figure 5: ATS automated drug screening platform
3. Protein crystallization : high-throughput, micro-system protein crystallization; reduce seed matrix effect, improve crystallization success rate
The protein crystallization process can be divided into three stages of nucleation, growth and stop growth. The formation of crystal nucleus is the starting point and key point of protein crystal growth, and the success rate of crystallization can be improved by seed crystal. The seeding process introduces the effect of the additional buffer solution on the crystallization conditions. For the first time, Roche scientists applied sonic pipetting techniques to the screening of high-throughput protein crystals. They reduced the volume of seed crystals to 15 nl by ATS, reducing the effect of buffer on the crystallization environment [2] .
As shown in Fig. 6, the blue histogram is seeded by ATS, and the orange and yellow bar graphs are the control group without seed crystals. Six protein samples were counted, and the number of crystal droplets in 96 droplets per protein. In another article [3] , Roche scientists used ATS to transfer protein solutions and various crystallization reagents to achieve 30nl system crystallization (Figure 7) and high-throughput crystallization conditions.
Figure 6: ATS seed crystal data
Figure 7: Crystallization of lysozyme 30nl system based on ATS pipetting
4. High-throughput mass spectrometry : ATS high-throughput mass spectrometry sample spotting, 100 times more efficient
The National Key Laboratory of Berkeley, USA, established a high-throughput mass spectrometry method based on ATS in order to optimize the microbial decomposition of polysaccharides and the production conditions of biofuels. ATS Transfer 1 nl of medium from a 384-well plate to a mass spectrometric sample metal plate (Figure 8) and perform mass spectrometry analysis of the microbial metabolites. The ATS completes a sample matrix with a point spacing of 450um and 288 points in 2 minutes. This method is more than 100 times more efficient than the manual method [4] .
Figure 8: High-throughput mass spectrometry sample spotting with ATS
To date, the world's top 10 pharmaceutical companies and many well-known research institutions have adopted ATS sonic pipetting technology for high-throughput drug screening, structural biology, molecular diagnostics, synthetic biology, precision medicine and other fields.
references:
[1] High-Throughput RT-PCR for small-molecule screening assays. Curr Protoc Chem Biol. ; 4(1): 49–63. doi:10.1002/9780470559277.ch110204.
[2] Acoustic matrix microseeding: improving protein crystalgrowth with minimal chemical bias
Acta Cryst. (2010). D66, 568–576
[3] Nanolitre-scale crystallization using acousticliquid-transfer technology. Acta Cryst. (2012). D68, 893–900
[4] High throughput nanostructure-initiator mass spectrometry screening of microbial growth conditions for maximalβ-glucosidase production. Frontiers in Microbiology.December 2013 | Volume 4 | Article 365 |
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