Multi-Modal Fluorescence

We developed a multimodal full-field optical coherence tomography (FF-OCT) and fluorescence microscope. Our system can simultaneously acquire FF-OCT and structured illumination microscopy images. Dynamic parallel evolution of tissue microstructures and biochemical environments can be visualized. We use high numerical aperture objectives to optimize the combination of the two modalities. We imaged the propagation of mechanical waves initiated by calcium waves in a heart wall to illustrate the interest of simultaneous recording of mechanical and biochemical information

Experimental setup with Green Fluorescent Protein configuration

Multimodal SIM/dynamic FF-OCT of a macaque retina. (a–d) Show the ganglion cell layer of a fresh macaque retina revealed by three different modalities acquired simultaneously with our microscope.

(a) The FF-OCT image and signal from ganglion cells are masked by collagen signal. A sparse
labeling of ganglion cells has been performed on this retina, and the corresponding SIM image is displayed in (b). (c) The same field of view imaged with dynamic FF-OCT, which removes the signal from the static collagen fibers to reveal several cells. The D-FF-OCT also reveals intracellular details. Finally,

(d) is the overlay between the dynamic FF-OCT image and the SIM image, revealing a perfect overlap between one of the ganglion cells and one cell revealed by dynamic FF-OCT. All the scale bars represent 35 μm.

O. Thouvenin, M. Fink, and C. Boccara, “Dynamic multimodal full-field optical coherence tomography and fluorescence structured illumination microscopy,” Journal of Biomedical Optics, vol. 22, no. 2, p. 026004, Feb. 2017.