Quantitative Label-Free Higher Harmonic Generation Microscopy for 3D and 4D Analysis of Tissue Structure and Dynamics
"I used a label-free microscopy technique (HHGM) to visualize and analyze cells and the tissue matrix around them (ECM) in models of the bone disease OI, the connective tissue disease EDS and lung diseases. In this way, we unravel how these diseases work.
"HHGM delivers "direct pathology": cells and structures can be seen in fresh tissue within minutes – much faster than classic H&E staining. Old methods such as immune staining or electron microscopy are static and time-consuming; Living cell staining in turn brings harmful dyes and light damage, which can change cells and only allow short exposures.
"My goal: to further develop HHGM into dynamic four-dimensional (4D) images of living tissue, and to determine what is needed for this.
"We show that label-free HHGM is a good tool to investigate 3D cultures of connective tissue cells (fibroblasts) and to measure collagen defects in the matrix that arise due to OI and EDS gene mutations.
"It also appears that completely label-free 4DHHGM can record images of elastin and collagen, contacts between cells and cell movement in fresh, precisely cut lung sections (PCLS) from humans – recorded as videos over a period of no less than five days."
More information on the thesis to follow.