CLSM reconstruction of methanotrophic bacteria in embryo tissue of deep-sea sponge H. (S.) methanophila, MTC849 probe

We double-labelled MTC849 (5` – CGTTAGCTCCACCACTAAG – 3) with Atto594 dye (Biomers, Ulm, Germany), and hybridized it with sponge tissue in 20% formamide buffer using standard protocol (Duperron <i>et al.</i>, 2008). Photomicrographs were acquired with confocal laser-scanning microscope (LSM 780, Carl Zeiss, Germany). Three channels were used - 594, DAPI and 488 (autofluoresence). 3D representation of image stacks was reconstructed with Fiji 3D project option (interpolated)..