Coherence Controlled Holographic Microscopy for Investigation of Cancer Cells Reactions in Turbid Microenvironment of Biologically Active Phospholipids

Coherence-Controlled Holographic Microscope (CCHM) provides quantitative phase imaging and in low coherence mode possesses pronounced Coherence Gate Effect (CGE). CGE in CCHM time-lapse mode offers the possibility of registration of cancer cells reactions even in turbid microenvironment. Thus the influence on cells of the substance that generates the turbidity as well as its cooperation with other added agents can be evaluated.

Emulsion of egg yolk phospholipids were shown to inhibit development of experimentally induced tumours in vivo as well as kill tumour cells in vitro (Karafiat et al., Folia Biol (Praha) 2014;60(5):220-7). However, the details of train of cellular events leading to cells death in such cytopathic turbid emulsion remained unnoticed. In addition biologically active phospholipids are getting into focus of anti-cancer research.

Therefore we decided to prove the expectations and in a model situation to test the CCHM capacity of imaging cells through emulsion at the high compared to the low coherence of illumination.

MCF-7 cells: Phase images through turbid emulsion. Coherent illumination (left, laser, 10 frames averaged) and low coherent illumination (right, halogen lamp, single shot). Calibration bar in pg/µm².

After confirming the assumptions we started investigation of attached DLD1 carcinoma cells incubated in turbid bioactive phospholipids causing eventually cell death. Results of CCHM recordings indicated necrosis as the kind of cell death. Experiments are in progress to show if an addition of other anti-cancer agents could modify this process to more favorable anti-cancer outcome which is apoptosis.