Detection of structural changes based on Mie-scattering analyses of mouse fibroblast L929 cells before and after necrosis

Abstract

The aim of the presented work is to investigate the angle-resolved scattering characteristics of biological nano- and micro-scaled cell structures. The scattering results of cellular structures were compared to measurements of ideal spherical nano- and micro-particles. A monolayer of mouse fibroblasts L929 cells was cultivated in a Dulbecco's Modified Eagle Medium (DMEM) in a standard 24 well cell culture plate. The system allows an in situ measurement directly in the standard cell culture plate and a contaminant-free investigation of the viability of the cell cultures. Of particular interest was whether changes in the tumor characteristics occur in apoptosis or other cell-harming effects. Because of the size ratios between wavelength and the scattering particles, all observations were investigated using Mie scattering theory. A setup for reliable measurements was developed and the scattered angle dependent intensity obtained was compared with simulated scattering characteristics. A homemade supercontinuum (SC) light source was filtered by an optical bandpass filter with a central wavelength of 500 nm. The scattered portion of the pulsed SC light behind the sample was recorded in a time-resolved manner at defined angles. A specimen holder adapted to standard cell culture plates allows detection of scattered radiation at angles between ±80° without angle-dependent Fresnel reflection losses and a Snell’s law bending of the propagation direction. Finally, the system was tested to detect structural changes of mouse fibroblast L929 cells before and after poisoning the cells with the cell detergent Triton X100 and the data clearly shows changes in the scattering characteristics when the cells were destroyed.