Branch Darren W, Vreeland Erika C, McClain Jamie L, Murton Jaclyn K, James Conrad D, Achyuthan Komandoor E
Nano and Micro Sensors Department, Sandia National Laboratories, Albuquerque, NM 87185, USA.
Imagion Biosystems, Inc., Albuquerque, NM 87106, USA.
Micromachines (Basel). 2017 Jul 21;8(7):228. doi: 10.3390/mi8070228.
Miniature ultrasonic lysis for biological sample preparation is a promising technique for efficient and rapid extraction of nucleic acids and proteins from a wide variety of biological sources. Acoustic methods achieve rapid, unbiased, and efficacious disruption of cellular membranes while avoiding the use of harsh chemicals and enzymes, which interfere with detection assays. In this work, a miniature acoustic nucleic acid extraction system is presented. Using a miniature bulk acoustic wave (BAW) transducer array based on 36° Y-cut lithium niobate, acoustic waves were coupled into disposable laminate-based microfluidic cartridges. To verify the lysing effectiveness, the amount of liberated ATP and the cell viability were measured and compared to untreated samples. The relationship between input power, energy dose, flow-rate, and lysing efficiency were determined. DNA was purified on-chip using three approaches implemented in the cartridges: a silica-based sol-gel silica-bead filled microchannel, nucleic acid binding magnetic beads, and Nafion-coated electrodes. Using the lysing dose defined as ATP released per joule was 2.2× greater, releasing 6.1× more ATP for the miniature BAW array compared to a bench-top acoustic lysis system. An electric field-based nucleic acid purification approach using Nafion films yielded an extraction efficiency of 69.2% in 10 min for 50 µL samples.
用于生物样品制备的微型超声裂解是一种很有前景的技术,可从多种生物来源高效快速地提取核酸和蛋白质。声学方法能够快速、无偏差且有效地破坏细胞膜,同时避免使用会干扰检测分析的苛刻化学物质和酶。在这项工作中,展示了一种微型声学核酸提取系统。使用基于36° Y切割铌酸锂的微型体声波(BAW)换能器阵列,将声波耦合到基于一次性层压材料的微流控芯片中。为了验证裂解效果,测量了释放的ATP量和细胞活力,并与未处理的样品进行比较。确定了输入功率、能量剂量、流速和裂解效率之间的关系。使用芯片中实现的三种方法在芯片上纯化DNA:基于二氧化硅的溶胶 - 凝胶填充有二氧化硅珠的微通道、核酸结合磁珠和涂有Nafion的电极。与台式声学裂解系统相比,使用每焦耳释放的ATP定义的裂解剂量,微型BAW阵列大2.2倍,释放的ATP多6.1倍。对于50 μL样品,使用基于Nafion膜的电场核酸纯化方法在10分钟内提取效率为69.2%。