Zhou Xuechang, Lau Lana, Lam Wendy Wai Ling, Au Shannon Wing Ngor, Zheng Bo
Department of Chemistry, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China.
Anal Chem. 2007 Jul 1;79(13):4924-30. doi: 10.1021/ac070306p. Epub 2007 Jun 5.
This paper describes a method of dispensing a nanoliter volume of liquid into arrays of microwells through degassed poly(dimethylsiloxane) (PDMS) microchannels. In this method, the PDMS microchannels were reversibly bound to arrays of microwells. The PDMS elastomer was predegassed and served as an internal vacuum pumping source. Various aqueous solutions were infused into arrays of microwells through the reversibly sealed PDMS microchannels. Microwells fabricated in PDMS, poly(methyl methacrylate) (PMMA), and glass were all compatible with this dispensing method. By removing the PDMS microchannels, arrays of droplets confined in the microwells were obtained. Multiplex reaction and screening at the nanoliter scale were carried out by binding two such arrays of microwells to form microchambers. We applied this method to screening the crystallization conditions of four known proteins. Long-term incubation of over 2 months was achieved by employing glass microwells. An unknown protein was then crystallized using the screening method in microwells. The crystals with sufficient size were harvested from the reversibly bound microwells. X-ray diffraction with a resolution of 3.1 Angstrom was obtained.
本文描述了一种通过脱气的聚二甲基硅氧烷（PDMS）微通道将纳升体积的液体分配到微孔阵列中的方法。在该方法中，PDMS微通道与微孔阵列可逆结合。PDMS弹性体预先脱气并用作内部真空泵浦源。各种水溶液通过可逆密封的PDMS微通道注入到微孔阵列中。在PDMS、聚甲基丙烯酸甲酯（PMMA）和玻璃中制造的微孔都与这种分配方法兼容。通过移除PDMS微通道，获得了限制在微孔中的液滴阵列。通过将两个这样的微孔阵列结合形成微室，在纳升尺度上进行多重反应和筛选。我们将此方法应用于筛选四种已知蛋白质的结晶条件。通过使用玻璃微孔实现了超过2个月的长期孵育。然后使用微孔中的筛选方法使一种未知蛋白质结晶。从可逆结合的微孔中收获了足够大小的晶体。获得了分辨率为3.1埃的X射线衍射结果。
