The Preliminary Study Of Microfluidic Immunoassay With Single-Molecules Detection

The thesis includes three parts:Ⅰ.The preliminary study of the conditions of microfluidic immunoassay by single molecules detection.ⅡThe optimization and choice of the conditions of microfluidic immunoassay by single molecules detection.Ⅲ.Detecting conditions of Cancer antigen 125 single molecules.In chapter one of this dissertation,a brief review of microfluidic,immunoassay analysis and single molecules detection was made.In this part,60 references have been referred.Then the preliminary study of the conditions of microfluidic immunoassay by single molecules detection was made.Several parameters including modification of PDMS,silanization time,driving voltage,buffer system were optimized.The following conditions were suitable for microfluidic immunoassay: modified PDMS slab with nascent oxygen for 2 days;the glass coverslip was silanized with 3%GOPS for 24 h;buffer washing conditions:injection voltage,1000 V,injection time,20 min;blocking conditions:injection voltage,1000 V,injection time 10 s,quiet time 50 s,repeat this precess:80 times;incubation conditions: injection voltage,1000 V,injection time 10 s,quiet time 50 s,repeat this precess:80 times.In chapter two of this dissertation,we made a further investigation about the microfluidic immnunoassay conditions on the base of chapter one.The diameter of the reservoirs,the fabrication and dimension of the microchannels were changed.All channels on the～5-mm thickness PDMS slab with a length of 4 cm and a width of 2 cm were～150μm wide and～20μm deep.On this chip we investigated the washing conditions,blocking time and incubation time etc..In chapter three of this dissertation,we investigate the conditions of sandwich immunoassay with single molecule detection on the microfluidic chip.The following conditions were suitable for microfluidic immunoassay to detect CA 125:...