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Microfluidic-Technology-Based Multiplexed Protein And Nucleic Acid Blotting Analysis

Posted on:2013-01-30Degree:MasterType:Thesis
Country:ChinaCandidate:S HeFull Text:PDF
GTID:2230330362473795Subject:Biomedical engineering
Abstract/Summary:PDF Full Text Request
Blotting is a crucial technique in many biological branches and has a very widerange of applications. However, some intrinsic problems have not been resolved withdevelopment in so many years. Along with the rapid accumulation of the knowledgefrom cell biology, molecular biology and proteomics, there is an increasingly need forobtaining more information about cellular signaling pathways and protein-proteininteractions with very limited samples. This is a grand challenge for the traditionalblotting technique because the traditional blotting technique has a very limitedprocessing throughput and is very cumbersome in procedures, far from satisfying theneed for modern biomedical diagnosis.Microfluidics is a newly developed technique in recent years. It utilizes micro-scalechannels and chambers to process the fluids and analytes and has the advantage of highthroughput, low cost and easy manipulation, which is the perfect compensation for thedrawbacks of traditional blotting technique. We have resolved several problems:We used soft lithography to fabricate the microfluidic chips and tune relativeparameters;We used three different methods to enhance the adhesion condition between PDMSand PVDF;We used the microfluidic chip to improve the performance of traditional blottingprocess, including multiplexed detection in multiple samples, internal molecular weightmarkers, simultaneous determination of antibody,antigen titration. We also investigatedthe influence of incubation time to the results and explored how to differentiate theneighboring bands using different channels;We also provided some possible explanations to the difference between the microfluidicsystem and the bulk systems;We applied this analytical system to the nucleic acid blotting and also proved itscapability.With these work, I established an analytical system that is capable of processingblotting samples and discovered some unique features. The components and principlesthat I applied are very simple so that it could possibly be commercialized to promote thebasic biological research in a very wide range.
Keywords/Search Tags:Biomedical Diagnosis, Microfluidics, Western Blot, Southern Blot
PDF Full Text Request
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