Lightsheet microscopy shows highly fluorescent dots in zebrafish embryos injected with both wt and mutated hCRYAB and hMYOT. Whole-mount immunofluorescence analysis with specific myotilin and αb-crystallin antibodies was performed at 48 hpf, and images of muscle fibers in the trunk region were acquired with a Lightsheet Z1 microscope (Zeiss International, Oberkochen, Germany). Images are Maximum Intensity Projections of selected planes for zebrafish wildtype embryos not injected (A,D,D′) or injected with hMYOT WT (B,B′), hMYOT S95I (C,C′,C″), hCRYAB WT (E,E′), hCRYAB G154S (F,F′). Total number of embryos analyzed for each experimental condition n = 3. Three independent experimental analyses have been performed. Panels (B′,C′,C″,D′,E′,F′) represent zoomed images from panels (B,C,D,E,F), respectively. Scale bar is 50 μm for (A–F) panels and 20 μm for (B′,C′,C″,D′,E′,F′) panels. Highly fluorescent dots variable in shape and size (black and white arrows), extended myotendinous junction (grey arrow in (F)) and deranged muscle fibers (large gray head arrow in (F)) are indicated.
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