Hydraulically coupled microejection technique for precise local solution delivery in tissues.
Hydraulically coupled microejection technique for precise local solution delivery in tissues.
复制标题
液压耦合微喷射技术，用于组织中精确的局部溶液输送。
DOI：
10.1016/j.jneumeth.2006.01.007
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发表时间：
2006
影响因子：
3
通讯作者：
Toney,GlennM
中科院分区：
文献类型：
--
作者：
Pakhomov,AndreiG;Semenov,Iurii;Brenner,Robert;Toney,GlennM
采用液压耦合体积微喷射（HCVM）来克服在离体脑切片和其他组织中常规压力喷射药物的已知缺点。对于HCVM，通过低顺应性的液体填充管将填充溶液的玻璃微量移液管连接到电机驱动的液体填充注射器，并且密封系统，不留下间隙或气泡。从微量移液器排出的体积与注射器柱塞移动所排出的体积相同（其不太受微量移液器尖端的直径或其被组织碎片堵塞的影响），因此可以精确地控制排出速率和持续时间。HCVM性能的特点是荧光成像的喷射染料，其组合与红外/微分干涉对比（IR/DIC）成像在脑切片，并通过并发膜片钳记录的喷射药物在个别神经元的影响。将不同体积的注射物注射到脑切片中（1.6- 400 nl）形成了一个球形，该球形短暂地移位了与微量移液管尖端相邻的组织。喷射染料的径向渗透与同时喷射药物的电生理反应良好相关，至少在喷射后的短暂（100- 200 ms）间隔内。穿透距离可以通过增加喷射体积或喷射速率或两者来增加。然而，在较高的喷射体积（100- 400 nl）下，显著部分的喷射物被组织弹性“推出”切片，并且通过沿喷射移液管的回流沿着逸出到外部。HCVM的使用使我们能够实现浓度上升时间（10-90%）在20- 40毫秒的数量级。这些测量和其他测量表明了HCVM的主要能力和局限性，并为其实际使用提供了指导，包括一些潜在的独特应用。
Hydraulically coupled volume microejection (HCVM) was employed to overcome known drawbacks of conventional pressure ejection of drugs in isolated brain slices and other tissues. For HCVM, a solution-filled glass micropipette is connected to a motor-driven, liquid-filled syringe by low-compliance, liquid-filled tubing, and the system is sealed leaving no gaps or air bubbles. The volume ejected from the micropipette is the same as the volume displaced by movement of the syringe plunger (it is not much influenced by the diameter of the micropipette tip or its clogging by tissue debris), so the ejection rate and duration can be precisely controlled. The HCVM performance was characterized by fluorescent imaging of ejected dyes, its combination with infrared/differential interference contrast (IR/DIC) imaging in brain slices, and by concurrent patch-clamp recording of ejected drug effects in individual neurons. Ejection of varied volumes into a brain slice (1.6–400nl) formed a globular shape that transiently displaced the tissue adjacent to the micropipette tip. The radial penetration of the ejected dye correlated well with the electrophysiological responses to a concurrently ejected drug, at least for brief (100–200ms) intervals after the ejection. The penetration distance could be increased by increasing either the ejection volume, or the ejection rate, or both. However, at higher ejection volumes (100–400nl), a notable fraction of the ejectate was “pushed” out of the slice by tissue resiliency, and escaped to the outside by a back-flow along the ejection pipette. The use of HCVM enabled us to achieve a concentration rise time (10–90%) on the order of 20–40ms. These and other measurements demonstrated principal capabilities and limitations of HCVM and provided guidance for its practical use, including some potentially unique applications.