Journal: ACS Omega
Article Title: Analysis of Intracellular Fatty Acid Metabolism during Doxorubicin-Induced Senescence of MCF7 Cells Using Raman Imaging
doi: 10.1021/acsomega.5c09213
Figure Lengend Snippet: Visualization of arachidonic acid metabolism in senescent MCF7 cells using hyperspectral Raman imaging of deuterated arachidonic acid. (A) Raman spectrum of 5,6,8,9,11,12,14,15- d 8 arachidonic acid (AA- d 8 ) showing two modes of signature spectral shifts of (CC)D stretching in the biologically silent region at 2220 (minor peak) and 2254 cm –1 (major peak). (B) Intensity of the two (CC)D stretching peaks (2220 and 2254 cm –1 ) normalized to the intensity of the total lipid peak (CH 2 stretching, 2850 cm –1 ) measured by Raman spectroscopy at different time points during PTGS2-mediated metabolism of arachidonic acid in vitro ( n ≥ 15). (C) Visualization of the intracellular distribution of AA- d 8 in senescent MCF7 cells by hyperspectral Raman imaging. (D) Ratiometric heatmaps for visualization of the intracellular distribution of AA- d 8 in senescent MCF7 cells by hyperspectral Raman imaging. (E) Intensity of the two (CC)D stretching peaks (2220 and 2254 cm –1 ) normalized to the intensity of the total lipid peak (CH 2 stretching, 2850 cm –1 ) measured by hyperspectral Raman imaging in senescent MCF7 cells at different time points after the removal of PTGS2 (COX2) inhibitor (Cay-10404). (The standard deviation between replicates was plotted as error bars. Statistical significance was tested by the two-tailed Student’s t test assuming heteroscedastic distributions. *** p < 0.001, **** p < 0.0001.)
Article Snippet: Hyperspectral Raman imaging was done using an alpha 300 Ri system (WITec GmbH, Oxford Instruments) using the following parameters: objective: 40× air; laser: 532 nm; laser power: 60 mW; scan speed: 2 s/pixel; pixel size: 1 μm/pixel; detector grating: 600 mm –1 .
Techniques: Imaging, Raman Spectroscopy, In Vitro, Standard Deviation, Two Tailed Test