Purpose Molecular imaging with Tc-99m-methoxy-isobutyl-isonitrile (Tc-99m-MIBI, MIBI) has been used in the assessment of thyroid nodules (TNs) for more than two decades. Many studies showed that MIBI imaging is a suitable tool to rule-out malignancy when negative. However, relatively low specificity and accuracy have been described, thus, limiting its acceptance in clinical practice. Additionally, different technologies, protocols, and interpretation criteria are adopted accounting for heterogeneous data reported in the literature. Therefore, the present study was undertaken to assess the clinical use and methodology of MIBI imaging in patients with nodular thyroid disease in Europe. Methods A questionnaire was sent to 12 European centers of Nuclear Medicine. The questionnaire encompassed ultrasound (US) and fine-needle aspiration cytology (FNAC) procedures and their evaluation as well scintigraphy imaging indications, technical procedures, and interpretation criteria of MIBI imaging. Results The survey showed a good agreement of different centers in approaching TNs by TSH measurement, US evaluation and Tc-99m-pertechnetate thyroid scintigraphy. MIBI imaging is mainly used to assess TNs with inconclusive/indeterminate cytological findings and selection of target nodule(s) for FNAC in patients with multi-nodular goiter. Technical procedures adopted in different centers are globally comparable and the recorded differences are unlikely to impact clinical results. However, as the main result of the present study, substantial differences were found in interpretation criteria adopted in different centers. Conclusions Our survey supports the urgent need of standardized interpretation criteria of thyroid MIBI imaging in order to improve its diagnostic performance and make results comparable in clinical practice.