AbstractSemiconductor quantum dots (QD) possess unique optical and electric properties like size‐tunable light emission, narrow emission range, high brightness and photostability. Recent research advances have minimized the toxicity of QDs and they are successfully used in in vitro and in vivo imaging. Encapsulation of QDs into polymeric nanoparticles and linking them with targeting ligands enabled the detection of tumors and cancer cells in vivo. QD‐antibody conjugates were successfully used in monitoring and diagnosis of HIV and myocardial infarction. Application of near infrared (NIR) QDs was found to minimize the absorption and scattering of light by native tissues thus rendering them suitable in deep tissue analysis. Aggregation and endosomal sequestration of QDs pose major challenges for the effective delivery of QDs to the cell cytosol. Toxicity minimization and effective delivery strategies may further increase their suitability for utilization in disease diagnosis. New synthesis of QDs may provide new types of bioconjugates of QDs to biomolecules, which leads to a variety of applications to many challenged research areas. QDs with narrow emission wavelength ranges are very suitable for monitoring multiple cellular targets simultaneously, and still remain the best known probes for imaging as an alternative to traditional fluorophores in disease diagnosis.