Features of Construction of the Fluorescent Microscope for the Study of Epithelial-Mesenchymal Transition of Cells in Vitro

Abstract

An optical luminescent microscope has been developed to monitor the behavior of tumor cells that were modified by GFP and RFP fluorescent proteins and to monitor the formation of metastases in biochips. Images of cells in the light of GFP and RFP fluorescence proteins, as well as in the photography mode, are recorded in the microscope by an noncooled CMOS matrix at the excitation of luminescence by blue and green LEDs and illumination of cells with white LEDs, respectively. The obtained fluorescent images and photos in white light are recorded by the matrix in the same coordinate system and do not require additional processing for combination. The power of irradiation of cells with LEDs does not exceed 50 μW/mm² at an exposure time of 1 s, which is an order of magnitude lower than the radiation power causing photodestruction and phototoxicity of cells during long-term studies. Relative parameters of the optical channel of the luminescent microscope have been introduced that allow comparing the sensitivity of the recording system and the minimum power level of the radiation that excites luminescence for optical elements with different spectral characteristics.