Bioimaging is changing the field of sensory biology, especially for taxa that are lesser-known, rare, and logistically difficult to source. When integrated with traditional neurobiological approaches, developing an archival, digital repository of morphological images can offer the opportunity to improve our understanding of whole neural systems without the issues of surgical intervention and negate the risk of damage and artefactual interpretation. This review focuses on current approaches to bioimaging the peripheral (sense organs) and central (brain) nervous systems in extant fishes (cartilaginous and bony) and non-avian reptiles in situ. Magnetic resonance imaging (MRI), micro-computed tomography (μCT), both super-resolution track density imaging and diffusion tensor-based imaging, and a range of other new technological advances are presented, together with novel approaches in optimizing both contrast and resolution, for developing detailed neuroanatomical atlases and enhancing comparative analyses of museum specimens. For MRI, tissue preparation, including choice of fixative, impacts tissue MR responses, where both resolving power and signal-to-noise ratio improve as field strength increases. Time in fixative, concentration of contrast agent, and duration of immersion in the contrast agent can also significantly affect relaxation times, and thus image quality. For μCT, the use of contrast-enhancing stains (iodine-, non-iodine-, or nanoparticle-based) is critical, where the type of fixative used, and the concentration of stain and duration of staining time often require species-specific optimization. Advanced reconstruction algorithms to reduce noise and artifacts and post-processing techniques, such as deconvolution and filtering, are now being used to improve image quality and resolution.