Synergy neo 2 hybrid multi mode microplate reader fluorescencespectrometer

Excitation–emission
matrices of A375 and HaCaT cell lines treated with oxaliplatin were
acquired by a Synergy Neo 2 Hybrid Multi-Mode Microplate Reader fluorescence
spectrometer (BioTek Instruments, Inc., Winooski, VT, USA) using a
Corning 96-well High Content Imaging Plate (Corning 4517, Corning,
Inc., Corning, NY, USA). A hand-written measurement protocol in which
subsequent emission spectra were recorded at decreasing excitation
wavelengths from 500 to 250 nm was used for the EEM fluorescence data
collection. The data interval in excitation mode was 10 nm. The range
of the recorded emission spectra depended on the excitation wavelength
at which the spectrum was acquired. For λ_ex in a
range of 290–500 nm, the emission was recorded from λ_em = λ_ex + 20 nm to 650 nm, whereas for λ_ex < 290 nm, the emission was measured in the range of 300–650
nm. The resolution of all emission spectra was 5 nm. All fluorescence
measurements were performed at 37 °C, using bottom optics. Each
sample, treated with oxaliplatin at various concentration levels,
was measured in 12 replicates.

The fluorescence measurements of UV irradiated cell culture were performed by Synergy™ Neo 2 Hybrid Multi-Mode Microplate Reader fluorescence spectrometer (BioTek Instruments, Inc., Winooski, VT, USA). The acquisition of excitation–emission matrices (EEMs) was realized using a hand-written measurement protocol in which subsequent emission spectra were recorded at decreasing excitation wavelength from 500 nm to 250 nm (with 10 nm interval). The range of the recorded emission spectra depended on the excitation wavelength at which the spectrum was measured. Thus, for λ_ex in a range of 290–500 nm, the emission was recorded from λ_em = λ_ex + 20 nm to 650 nm. In the case of λ_ex <290 nm, the emission was recorded over the spectral range of 300–650 nm (Fig. 3). This method of EEM spectra acquisition allowed to avoid Rayleigh and Raman signals in the obtained spectra. The resolution of all emission spectra was 5 nm. All fluorescence measurements were carried out at 37 °C, using bottom optics and Corning® 96-well High Content Imaging Plate (Corning 4517, Corning, Inc., Corning, NY, USA). Each sample, treated by UV radiation of different time duration (0, 10, 20, 40, 50, 60 min), was measured in 12 replicates.