Figure 4
TPEF and THG images of Phycomyces blakesleeanus exponential growth phase hyphae in standard liquid medium show that the predominant source of spot wise THG signal are lipid droplets. (a) Merged autoTPEF and THG images of same unlabeled live hypha showing that there is no overlap of autoTPEF and THG signal. The average laser power at sample plane was 28 mW at 1040 nm (for THG) and 3.4 mW at 730 nm (autoTPEF). (b) In vivo colocalization of stained LDs imaged by TPEF and LDs imaged by THG modality. Average laser power at sample plane was 32 mW for both THG and TPEF at 1040 nm. Pearson’s correlation coefficient Rtotal = 0.844 (ImageJ, The Colocalization Threshold plugin). All images were taken with Zeiss 40 × 1.3 oil objective lens.
