Electromagnetic field interactions with micro channels, particles and cells: Application to advanced cytometry
Abstract
This thesis involves a study of the interaction of laser beams with micro channels and micro particles/cells using the electromagnetic field approach. This problem is relevant to the next generation cytometry, in particular to model based design of flow cytometers. The field approach is applied to study light scatter from particles/cells and also internal and scattered fields of cylindrical micro channels that are important for optical interrogation of particles and cells flowing through. Though current flow cytometers use qualitative fluorescence measurements for biological analysis, other viable optical interrogation techniques like light scatter, quantitative fluorescence and Coherent anti-stokes Raman scatter (CARS) are being studied for application to flow cytometry. The light scatter from particles and cells in a flow cytometer has been studied with the objective of extracting useful information about the particles using scatter measurements. First, the correlation between the size of particles and the current forward scatter measurements was both analytically modeled and experimentally determined. These results indicated that integrated scatter measurements currently used in flow cytometry (forward and side scatter) cannot be used to unambiguously estimate size, shape or refractive index of particles for classification. It is shown that multi-angle scatter measurements can be used to classify micro spheres of different sizes/refractive indices and different bacteria species, provided the scatter measurements are designed based on numerical scatter models. The numerical scatter models were then also used to do a preliminary study of correlation of scatter with internal structure of simple cells like stem cells. A few multivariate statistical methods have been applied for the classification of such particles in flow cytometry using scatter and multi-spectral fluorescence measurements. Typically the micro channels used in flow cytometry have square or circular cross sections and the channels as such are open cuboids or circular cylinders. The next generation flow cytometry would involve measurements of light scatter at multiple angles and multiple laser beams incident at different angles (for CARS) and it would be advantageous to use cylindrical channels. Hence the problem of interaction of shaped laser beams and cylindrical micro channels is solved in this thesis.
Degree
Ph.D.
Advisors
Robinson, Purdue University.
Subject Area
Biomedical research|Electromagnetism
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