Date of Award

Summer 8-16-2019

Level of Access Assigned by Author

Open-Access Thesis

Degree Name

Master of Science (MS)


Biological Engineering


Karissa Tilbury

Second Committee Member

Michael Mason

Third Committee Member

Paul Millard

Additional Committee Members

Robert Bayer


Two-photon microscopy (TPM) is a powerful, versatile imaging modality for the study of biological systems. This thesis overviews the relevant physics involved in TPM, design considerations and process of constructing a home-built, two-photon microscope, and provides a set of procedures to operate the system. Furthermore, this work explores several applications of TPM through the study of single-cell metabolism and imaging the cellular-material interface. Explored in particular depth was the imaging of cellulose nanofiber (CNF) materials, with the goal of understanding the three-dimensional nature of fibroblast cell growth when embedded within the materials. This work uncovered several optical properties of CNF, and resulted in the creation of a novel method for imaging three-dimensional cellular growth into CNF materials; resolving fine material and biological structures without perturbing the native environment.