Microarrays for High Throughput Analysis of Cellular Heterogeneity
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The importance of cellular heterogeneity within cell populations has been increasingly realized in recent years. As a result, there has been a focus on developing methods for single cell analysis. µCP represents a cheap, simple and versatile method for producing microarrays across a substrate surfaces. The microarrays can be utilized for cell immobilization, which can be coupled to microscopy techniques. There is also an increased interest in using AFM for cell studies, which requires immobilization The focus of this thesis was to develop a method for µCP of PLL to facilitate immobilization of S. cerevisiae on a glass substrate. The idea is to create a pattern that enables immobilization of single cells of S. cerevisiae to achieve SCA. A photolithography procedure was developed to create a master mould for replica moulding. An even film thickness was achieved, but there were persistent problems with edge bead and cracks in the resist due to thermal stress. Replica moulding was used to make PDMS stamps on the created master mould. AFM analysis showed irregularities in the stamps corresponding to the cracks observed in the resist, which resulted in poor conformal contact between the stamp and the substrate surface. The parameters of the replica moulding method were verified using a pre-made master mould to create stamps. This showed that the process was suitable for creating PDMS stamps with micrometre scale features. The stamps were used to demonstrate that PDMS stamps could be used to print PLL in a micrometre scale pattern. It was also demonstrated that S. cerevisiae could be immobilized on glass substrates functionalized with PLL.