The ultrasound contrast agents currently used in clinics are microbubbles with a large particle size and short circulation time, and their approved clinical applications are limited to endovascular diagnosis and therapy only. The development of ultrasound-responsive nanodroplets (NDs) provides a new approach for extravascular diagnosis and therapy, especially for molecular imaging and targeted therapy of tumors. The NDs with a nano-scaled particle size and a liquid core can maintain their shape and initial diameter during injection, enhancing their EPR effects and facilitating the accumulation of NDs at the tumor site. When exposed to ultrasound, NDs can vaporize and exhibit contrast enhancement at the sites of interest. In addition, the destruction of microbubbles can provide a driving force to facilitate the release of drugs or genes from the microbubbles into target cells, allowing the NDs to act as drug carriers. The development of ultrasound-responsive NDs has shown rapid progress in recent years, while a variety of NDs with excellent properties have been fabricated for targeted diagnosis and drug delivery. In this article, the development of ultrasound-responsive NDs was reviewed in terms of their structure, phase transition properties, and applications in targeted tumor diagnosis and therapy.