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The entrapment and characterization of single spermatozoa in a microfluidic system

Berendsen, Jorien (2014) The entrapment and characterization of single spermatozoa in a microfluidic system.

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Abstract:This research describes the entrapment of spermatozoa in a micro uidic device and the analysis of single spermatozoa by uorescence and impedance spectroscopy. This micro uidic device is made from PDMS and consists of two main channels, which are con- nected by small side channels. These side channels act as small traps when a pressure difference is obtained over the main channels. This pressure difference was caused by a difference in ow rate. Three different trap heights of 1, 1.5 and 2 mm were tested, of which the 1 mm traps showed a single cell trapping capacity of 43%. This chip showed the least tendency to trap multiple sper- matozoa, which is an advantage since multiple trapping is undesirable for impedance measurements. Spermatozoa were analysed with uorescent stainings. Viability staining was performed on and off chip. With viability staining on chip, the viability of a specific cell could be determined. FISH staining of X and Y chromosomes was performed, after which several FISH staining experiments were performed on spermatozoa on chip. The staining results were not in uenced by the presence of microelectrodes in the chip. The presence of a spermatozoon in the trap could also be detected through impedance spectroscopy. It was seen that spermatozoa caused a drop in impedance at the resistive plateau. This drop in impedance indicates that the interior of the cell was measured. Because the interior of spermatozoa mainly consists of DNA, it might be possible to measure the amount of DNA. Therefore, viability and FISH staining on chip can be combined with impedance measurements on chip, to investigate the impedance differences of viable and dead spermatozoa and of X and Y bearing spermatozoa.
Item Type:Essay (Bachelor)
Faculty:TNW: Science and Technology
Subject:50 technical science in general
Programme:Advanced Technology BSc (50002)
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