<p>Fluorescence intensity line-profile analysis of metabolic enzyme staining relative to nuclear markers in <i>T. cruzi.</i></p>

HK, CS, and GK were detected with Alexa Fluor 488-conjugated secondary antibody, H2B.V-Myc was detected with Alexa Fluor 555-conjugated secondary antibody, and DNA was stained with DAPI. Images were acquired using an Olympus BX51 fluorescence microscope controlled by Olympus cell^R software.

White arrows indicate the linear regions of interest drawn across the parasite body and nuclear region. The same linear ROI was applied to the Alexa Fluor 488, Alexa Fluor 555, and DAPI channels, and fluorescence intensity values were extracted along the ROI using Olympus cell^R. Intensity values were normalized and plotted as line profiles in GraphPad Prism to compare the spatial distribution of the three fluorescence signals.</p> <p>(TIF)</p>