||The photonic nanojet is a non-resonance focusing phenomenon with high intensity and narrow spot that can serve as a powerful biosensor for in vivo detection of red blood cells, micro-organisms, and tumor cells in blood. In this study, we first demonstrated photonic nanojet modulation by utilizing a spider-silk-based metal–dielectric dome microlens. A cellar spider was employed in extracting the silk fiber, which possesses a liquid-collecting ability to form a dielectric dome microlens. The metal casing on the surface of the dielectric dome was coated by using a glancing angle deposition technique. Due to the nature of surface plasmon polaritons, the characteristics of photonic nanojets are strongly modulated by different metal casings. Numerical and experimental results showed that the intensity of the photonic nanojet was increased by a factor of three for the gold-coated dome microlens due to surface plasmon resonance. The spider-silk-based metal-dielectric dome microlens could be used to scan a biological target for large-area imaging with a conventional optical microscope.