Resolution Enhancement in Two-photon Microscopy by Applying Structured Line Illumination

• Main Authors: Chi-deng Lin, 林祺登
• Other Authors: Szu-yu Chen
• Format: Others
• Language: zh-TW
• Published: 2015
• Online Access: http://ndltd.ncl.edu.tw/handle/01902156988926275896

碩士 === 國立中央大學 === 光電科學與工程學系 === 103 === The aim of this study is to enhance the spatial resolution in two-photon microscopy (TPM) by applying a line-shaped structured illumination. Since structured illumination microscopy (SIM) expands the effective frequency spectrum space twice the size bigger of a traditional microscopy’s, its spatial resolution is therefore two times better. Combining it with TPM’s optical sectioning ability, which is due to the relation between the emission and excitation intensity, we can increase the vertical spatial resolution, and also raise the image’s spatial resolution up nearly three times better. Therefore, utilizing our system not only improves the lateral resolution, but also provides optical sectioning ability. This study is separated into simulation and experimental parts. In the simulation part, by using the parameters shown in the spec of the experiment setup, we simulate the whole process of a TPM image being excited by a structured line illumination, and then use the results to reconstruct the image via algorithm, in which, we were able to come up with a improve factor of 2.57. Later in the experimental part, by using quantum dot nanospheres as our samples for lateral resolution measurement purposes, and exciting two-photon fluorescence while applying structured line illumination, we successfully reconstructed the image via algorithm with a lateral resolution improve factor of 1.7. Finally, we compare the experiment results with the simulation, and discuss its advantages.