OBJECTIVE: To determine the quality of samples for cytological analysis, to establish the correlation between the cytological image of Hashimoto's thyroiditis and the antibody titre level on the thyroid peroxidase (anti-TPO) and to establish the correlation between the cytological image of Hashimoto's thyroiditis and thyroglobulin antibody titre level (anti-Tg). STUDY DESIGN: Cross-sectional study. MATERIALS AND METHODS: The study included 66 participants with Hashimoto's thyroiditis, who underwent cytological analysis of thyroid gland fragments obtained by puncture by means of ultrasonography, in the period between January and June 2017, with determined levels of antibody titre on thyroid peroxidase and levels of antibody titre on thyroglobulin. Samples were dyed by May-Grünwald-Giemsa method and analysed by a light microscope. RESULTS: The cytological analysis was well prepared in 53% of the samples. A good quality sample was found in 44% of participants, and good colouring quality was found in 62% of samples. There are no significant differences in the distribution of participants based on anti-TPO and anti-Tg values. Higher antibody titre in case of Hashimoto's thyroiditis is more common in women. CONCLUSION: Targeted cytological thyroid gland puncture under ultrasonography control is a simple and reliable method in the diagnosis of Hashimoto's thyroiditis, but the precondition for quality cytological analysis is a well prepared, labelled and coloured sample of appropriate cellularity and availability of all cells for cytological analysis.