摘要
顯微成像橢偏儀（MIE）是一種光學技術，透過顯微鏡成像之下測量橢偏參數Psi（Ψ）和Delta（Δ）。這種技術公認的優勢在於非接觸、非破壞性和非侵入性。因此，它可以保持原本的條件之下檢測生物樣本的特性。.
在這項工作的第一部分，我使用MIE來測量乾燥的變形鏈球菌（變形鏈球菌）細胞在玻璃底物上的光學響應。使用具有單反射、雙反射和多重反射等三種不同理論模型分析500nm，600nm和700nm處的Ψ和Δ圖像，以獲得光學常數和高度分佈。經過檢測獲得變形鏈球菌的光學常數圖像之後，將單反射分析、雙反射或多重反射分析進行比較，顯示出不同的觀點。此外，通過原子力顯微鏡（AFM）測量的厚度值與變形鏈球菌樣品的雙反射和多重反射分析所估計的高度分佈一致，這意味著雙反射和多重反射分析可提供的信息與通過單反射分析所獲得的信息互補。.
在第二部分，我使用MIE研究在Au膜上培養的變形鏈球菌細胞的表面拓撲變化，通過測量可見光範圍（λ= 490nm-710nm）中的波長（λ）的橢偏光譜。對於一系列不同的物鏡平面（焦平面附近）收集二和四個細胞鏈的橢圓偏振係數圖像Ψ，Δ，p和s偏振反射率（Ip和Is）和反射率差分圖像（RDI） ），λ= 600nm。獲得的結果證明，我們能夠識別小於1μm的細胞並觀察其在Ψ和Δ圖像中的繞射圖像。我們發現Δ光譜和圖像對物鏡平面（POP）的位置特別敏感，而變異型鏈球菌細胞的Ψ光譜和圖像則對POP不敏感。

Microscopic Imaging Ellipsometry (MIE) is an optical technique that uses an objective and sensing procedure to measure the ellipsometric parameters Psi (Ψ) and Delta (Δ) in the form of microscopic maps. This technique is well known for being non-contact, non-destructive, and non-invasive. Therefore it can be used to detect and characterize biological species without any impact.
In the first part of this work, I used MIE to measure the optical response of dried Streptococcus mutans (S. mutans) cells on a glass substrate. The Ψ and Δ images at 500nm, 600nm, and 700nm were analyzed using three different theoretical models with single-bounce, two-bounce, and multi-bounce light paths to obtain the optical constants and height distribution. The obtained images of the optical constants show different aspects when comparing the single-bounce analysis with the two-bounce or multi-bounce analysis in detecting S. mutans samples. Furthermore, the height distributions estimated by two-bounce and multi-bounce analysis of S. mutans samples were in agreement with the thickness values measured by atomic force microscopy (AFM), which implies that the two-bounce and multi-bounce analysis can provide information complementary to that obtained by single-bounce light path.
In the second part, I used MIE to study surface topology variation for S. mutans cells cultured on Au film by measuring the ellipsometry spectra for wavelengths (λ) in the visible range (λ= 490nm - 710nm). The ellipsometry characteristic images Ψ, Δ, p- and s- polarized reflectance (Ip and Is), and reflectance difference image (RDI) for a chain of two and four cells were collected for a series of different objective planes (near the focal plane) for λ=600nm. The obtained results show that we were able to identify cells smaller than 1µm and observe their diffraction patterns in Ψ and Δ images. We found that the Δ spectra and images are particularly sensitive to the position of objective planes (POP), while the Ψ spectra and images for S. mutans cells are rather insensitive to POP.

摘要………...…………………………………………………………..ii
Abstract…………………………………………………………………iii
Acknowledgements…………………………………………………v
Chapter 1 Introduction ………………………………………………1
1.1 Streptococcus Mutans…………………………………1
1.2biofilm formation by S. mutans……………………………1
1.3 Ellipsometry…………………………………………………3
1.4 History of ellipsometry ………………………………………3
Chapter 2 Motivation ……………………………………………5
Chapter 3 Microscopic Imaging Ellipsometry………………………8
3.1MIE-RCE set up…………………………………………………8
3.2Calibration of MIE-RCE ………………………………………12
Chapter 4 Methods and Materials …………………………………………………13
4.1Cell culture ………………………………………………13
4.2Cell sample preparation ……………………………………13
4.3Theory and numerical calculations …………………………14
4.3.1Single-bounce light path …………………………16
4.3.1.1 s – polarization …………………16
4.3.1.2 p – polarization ……………………18
4.3.2Two-bounce light path ……………21
4.3.3Multi-bounce light path…………24
Chapter 5 Results and Discussion…………………………………27
5.1MIE-RCE calibration for glass substrate and fitting model..27
5.2MIE-RCE calibration for Si substrate and Au film………29
5.3AFM results ………………………………………………………31
5.4MIE-RCE results of S.mutans on glass substrate ..…………32
5.4.1Single-bounce analysis ……………………………36
5.4.2Two-bounce analysis …………………………39
5.4.3Multi-bounce analysis……………………………42
5.5 MIE-RCE results of S.mutans on Au film …………………47
5.5.1MIE images for different objective planes of two
S. mutans cells ……………………………………………………48
5.5.2MIE images for different objective planes of four
S. mutans cells …………………………………..............................52
5.5.3Significance of delta image for identifying
submicron feature ……………………………………………………56
5.5.4MIE spectra for different objective planes of two and four S. mutans cells …………………………………………………57
Chapter 6 Conclusion and Future Work ………………………………………60
6.1Conclusion …………………………………………………60
6.2Future work ………………………………………………62
References ………………………………………………………63
Appendix …………………………………………………74

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