Method to analyze effects of low-level laser therapy on biological cells with a digital holographic microscope.

Low-level laser therapy (LLLT) is a therapeutic tool that uses the photobiochemical interaction between light and tissue. Its effectiveness is controversial due to a strong dependence on dosimetric parameters. In this work, we demonstrate that digital holographic microscopy is an effective label-free imaging technique to analyze the effects of LLLT on biological cells, and we propose the full methodology to create correct synthetic aperture phase maps for further extensive, highly accurate statistical analysis. The proposed methodology has been designed to provide a basis for many other biological experiments using quantitative phase imaging. We use SHSY-5Y and HaCaT cells irradiated with different doses of red light for the experiment. The analysis shows quantitative changes in cell dry mass density and the projected cell surface in response to different radiation doses.

Volumetric segmentation of biological cells and subcellular structures for optical diffraction tomography images.

Three-dimensional, quantitative imaging of biological cells and their internal structures performed by optical diffraction tomography (ODT) is an important part of biomedical research. However, conducting quantitative analysis of ODT images requires performing 3D segmentation with high accuracy, often unattainable with available segmentation methods. Therefore, in this work, we present a new semi-automatic method, called ODT-SAS, which combines several non-machine-learning techniques to segment cells and 2 types of their organelles: nucleoli and lipid structures (LS). ODT-SAS has been compared with Cellpose and slice-by-slice manual segmentation, respectively, in cell segmentation and organelles segmentation. The comparison shows superiority of ODT-SAS over Cellpose and reveals the potential of our technique in detecting cells, nucleoli and LS.

Influence of Yokukansan on the refractive index of neuroblastoma cells.

Yokukansan (YKS) is a traditional Japanese herbal medicine that is increasingly being studied for its effects on neurodegenerative diseases. In our study, we presented a novel methodology for a multimodal analysis of the effects of YKS on nerve cells. The measurements of 3D refractive index distribution and its changes performed by holographic tomography were supported with an investigation by Raman micro-spectroscopy and fluorescence microscopy to gather complementary morphological and chemical information about cells and YKS influence. It was shown that at the concentrations tested, YKS inhibits proliferation, possibly involving reactive oxygen species. Also substantial changes in the cell RI after few hours of YKS exposure were detected, followed by longer-term changes in cell lipid composition and chromatin state.