Label-free imaging of heme proteins with two-photon excited photothermal lens microscopy

Heme proteins, such as hemoglobins and cytochromes, play important roles in various biological processes. Here we employ the two-photon excited photothermal effect as a contrast mechanism to map heme proteins distribution. Particularly, both a thermal lens scheme and a high-frequency modulation are utilized to enhance the signal-to-noise ratio. We demonstrate label-free imaging of individual red blood cells, subcellular distribution of cytochromes in live mammalian cells, and the microvascular networks in mouse ear tissue and in a zebrafish gill.