A compact disk-like centrifugal microfluidic system for high-throughput nanoliter-scale protein crystallization screening.
A centrifuge-based microfluidic system has been developed that enables automated high-throughput and low-volume protein crystallizations. In this system, protein solution was automatically and accurately metered and dispensed into nanoliter-sized multiple reaction chambers, and it was mixed with various types of precipitants using a combination of capillary effect and centrifugal force. It has the advantages of simple fabrication, easy operation, and extremely low waste. To demonstrate the feasibility of this system, we constructed a chip containing 24 units and used it to perform lysozyme and cyan fluorescent protein (CyPet) crystallization trials. The results demonstrate that high-quality crystals can be grown and harvested from such a nanoliter-volume microfluidic system. Compared to other microfluidic technologies for protein crystallization, this microfluidic system allows zero waste, simple structure and convenient operation, which suggests that our microfluidic disk can be applied not only to protein crystallization, but also to the miniaturization of various biochemical reactions requiring precise nanoscale control.
Li
Gang
G
Nanotechnology Laboratory, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, People's Republic of China. gang_li@mail.sim.ac.cn
Chen
Qiang
Q
Li
Junjun
J
Hu
Xiaojian
X
Zhao
Jianlong
J
D000818:Animals; D002498:Centrifugation; D011232:Chemical Precipitation; D016250:Compact Disks; D003460:Crystallization / Q000191:economics / Q000295:instrumentation*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000302:isolation & purification; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000302:isolation & purification; D046210:Microfluidic Analytical Techniques; D009113:Muramidase / Q000737:chemistry / Q000302:isolation & purification; D055620:Optical Phenomena; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification; D012996:Solutions; D013997:Time Factors
C485182:Cyan Fluorescent Protein; D029968:Escherichia coli Proteins; D011506:Proteins; D012996:Solutions; D049452:Green Fluorescent Proteins; D009113:Muramidase