Fuhr G, Müller T, Baukloh V, Lucas K
Humboldt-Universität zu Berlin, Institut für Biologie, Germany.
Hum Reprod. 1998 Jan;13(1):136-41. doi: 10.1093/humrep/13.1.136.
We present a new touch-free technique for trapping, positioning and selecting human spermatozoa. This can be done in free solution (culture medium) by high-frequency electric fields. Ultramicroelectrodes fabricated by photo- and electron-beam lithography on quartz and glass substrates were used to create field cages or long field channels. If the conductivity of the external salt solution is higher than the average value of sperm cell conductivity, negative polarization and negative dielectrophoresis occur. As a result, the induced cell polarization leads to forces repelling spermatozoa from the electrodes towards the field minimum. Using four planar electrodes a field funnel can be formed in which an individual spermatozoon is retarded while swimming. The same can be done more effectively in three-dimensional cages created by an octopole electrode system. In these systems, rapidly swimming spermatozoa could be trapped for several seconds but some spermatozoa stop moving if exposed to field strengths of more than 500 V/cm at frequencies in the MHz range. However, in stripwise and interdigitated electrodes, rapidly swimming sperm cells could be very well positioned in front of a break-electrode by a combination of electric field trapping and field induced laminar fluid streaming. This technique can be applied to bring individual spermatozoa to a defined position for characterization followed by sampling with capillaries.
我们提出了一种用于捕获、定位和筛选人类精子的新型非接触技术。这可以通过在自由溶液(培养基)中利用高频电场来实现。通过光刻和电子束光刻技术在石英和玻璃基板上制造的超微电极被用于创建场笼或长场通道。如果外部盐溶液的电导率高于精子细胞电导率的平均值,就会发生负极化和负介电泳。结果,诱导的细胞极化会产生将精子从电极排斥向场强最小值的力。使用四个平面电极可以形成一个场漏斗,单个精子在游动时会在其中减速。在由八极电极系统创建的三维笼中可以更有效地实现同样的效果。在这些系统中,快速游动的精子可以被捕获几秒钟,但如果在兆赫兹范围内的频率下暴露于超过500 V/cm的场强,一些精子会停止游动。然而,在条形和叉指电极中,通过电场捕获和场诱导层流的组合,快速游动的精子细胞可以很好地定位在断路电极前。这项技术可用于将单个精子带到特定位置进行表征,随后用毛细管进行采样。