Fig. 3.

Fluorescence images of HeLa cells were captured after a 30 min incubation with 10 μM probe A and 10 μM of IR-780 dye. The green channel (I) was used to observe the visible fluorescence of probe A in the 425–525 nm range, while the red channel (II) captured near-infrared fluorescence from probe A (600–700 nm). The green channel employed Alexa Fluor 405 (excitation at 405 nm), and the red channel utilized Alexa Fluor 568 (excitation at 559 nm). The magenta channel (III) was used to detect near-infrared fluorescence from the mitochondria-specific dye (IR-780) in the 700–800 nm range, with Alexa Fluor 635 (excitation at 633 nm). Scale bars represent 20 μm. A PCC value of 0.929 was obtained for the channel II/IR 780 overlay.