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Biophysical methods underlie advances for analytical, diagnostic, and therapeutic applications in biology and biomedicine. Electromagnetic radiation as visible and infrared light allows the sensitive collection of data on the morphofunctional conditions of biological substrates, while biomechanical approaches can improve therapeutic protocols. A common denominator is the need to optimize operating protocols, data processing, and interpretation, to achieve reliable information for analysis, diagnosis, and therapy.

This reprint from the Special Issue "Development of New Optical Techniques and Methods for Basic Biology and Biomedical Applications" intends to further disseminate awareness on the ample potential of biophysical method applications in biology and biomedicine, represented by all its contributing papers. Fluorescence spectroscopy is used for basic research on melaninization processes or for the noninvasive diagnosis of vascular circulation, and imaging of fluorescing Hydra polyp movements promotes hydrodynamic studies. Near-IR imaging is used with exogenous agents to enhance cardiac injury detection, to improve nanomaterial intracellular localization aimed to optimize drug delivery and cancer photo-thermal therapy, to delineate internal structures in animal bodies or to detect thermal changes in sites causing pain. In turn, a biomechanical approach entails the optimization of vibrational stimulation to restore weak muscles. All these reports improve the reliability of the various procedures proposed, hoping to inspire novel investigations.