透過分離與沉降法之精子品質檢測晶片

Sperm Quality Assessment via Separation and Sedimentation in a Microfluidic Device

蔣存超(Tsun-Chao Chiang)

指導教授 ： 胡文聰

共同指導教授 ： 鍾德憲(De-Shien Jong)

國立臺灣大學/工學院/應用力學研究所/碩士(2012年)

https://doi.org/10.6342/NTU.2012.01562

摘要

近年來台灣的生育率不斷的下降，近乎達到人口零成長的現象，造成社會人口結構的老化，對於國家的競爭力影響甚巨。在社會壓力、環境、結婚年紀上升等因素之影響，有六分之一計畫生育之夫妻有不孕症的困擾，其中因男性精子狀況不佳占約一半的比例。
現今的精子品質檢測系統，大部分需要高等級的顯微鏡搭配精子計數器藉由操作員或是軟體來檢測精子的總量、泳動率、形狀。精子診斷當中以精子的濃度與泳動率為優先判別的二種指標，惟現今的微流檢測系統或是染色判別系統只能提供單一種指標，本研究為提供一套微流系統可同時的檢測出精子的總量與泳動率。本研究利用精子泳動的特性在微流道當中可以分離出泳動與非泳動的精子，藉由離心機將精子沉降到檢測的流道當中，分析精子堆積的體積來反推出精子的濃度與泳動率。本研究與台大醫院泌尿科合作，在樣本數n=10所得到的結果與標準的檢測系統(Makler chamber)相比近乎95%。藉以佐證本研究所設計的晶片與現今的檢測系統具有相當程度的準確性，且同時提供兩種重要的品質因子。未來目標將微流晶片製作到光碟上，利用光碟機離心與電腦影像讀取，可居家快速檢測並將資料上傳至雲端提供醫生診斷的依據，且此套系統無須顯微設備，可減少大量的檢測成本。

關鍵字

精子品質 ； 不孕症 ； 微流體晶片 ； 精子計數 ； 沉降作用

並列摘要

A large percentage of infertility is due to male factor. Total spermatozoon concentration and their motile percentage are vital criteria for evaluation, yet difficult due to complicated factors. This paper presents a simple microfluidic device to assess human sperm quality by quantifying both total and motile sperm counts. Semen sample was allowed to reach equilibrium whereby non-motile remain in the inlet reservoir and roughly half of motile sperms swim across a phaseguide barrier into an adjacent region. Then centrifugation results in two pellet areas representing total and motile sperm concentrations upon calculation of the corresponding occupied areas. Results show centrifuge at 1500g for 20 mins generated compact and robust pellets for concentration less than 〖10〗^7 sperm/ml, and revealed good correlation against Makler chamber with coefficient of determination R^2=0.97. Motile percentage in range of 10-70% was tested, encompassing the cutoff value of 40% stated by WHO standard, with good coefficient of determination of R2= 0.84. Comparison of results from Makler Chamber, sperm quality analyzer (SQA) and the sperm microfluidic disk revealed results from the sperm disk agrees with the Makler chamber without statistical significance. The sperm microfluidic disk analyzes both total and motile sperm concentrations in one spin, accurate and easy to use, which should enable sperm quality analysis without the aid of expensive microscope.

並列關鍵字

Sperm analysis ； Fertility ； Microfluidic device ； Cell counting ； Sedimentation

參考文獻

[38] A. Donev, I. Cisse, D. Sachs, E. Variano, F. H. Stillinger, R. Connelly, S. Torquato, and P. M. Chaikin, "Improving the density of jammed disordered packings using ellipsoids," Science, vol. 303, pp. 990-993, Feb 13 2004.

[1] P. Andolz, M. A. Bielsa, and J. Vila, "Evolution of semen quality in north-eastern spain: A study in 22,759 infertile men over a 36 year period," Human Reproduction, vol. 14, pp. 731-735, Mar 1999.

[3] J. Lackner, G. Schatzl, T. Waldhor, K. Resch, C. Kratzik, and M. Marberger, "Constant decline in sperm concentration in infertile males in an urban population: Experience over 18 years," Fertility and Sterility, vol. 84, pp. 1657-1661, Dec 2005.

[4] H. N. Highland, B. Kalaria, and L. B. George, "A study on the trend in semen parameters of males from ahmedabad city (gujarat), over a two-decade period," Studies on Ethno-Medicine, vol. 4, pp. 199-202, Dec 2010.

[5] H. W. G. Baker, "The fifth edition of the who manual on semen analysis," Australian & New Zealand Journal of Obstetrics & Gynaecology, vol. 50, pp. 28-28, Oct 2010.