In its development so far, this department has shown great interest in research, study, collection and presentation of the rich cultural heritage of the Negotino region. At the Department of Ethnology, there is a presentation of the material and spiritual culture of the settlements in the Municipality of Negotino and the city of Negotino, through numerous folk dresses, items that were used in the household and the agricultural culture prevailing in this region from the beginning of the 18th to the end of the 20th century. These include, part of which are exhibited, different copper jugs, baking tins, boilers, small copper jugs, distaffs/spindles, low round wooden dining tables, sickles, gourds, pitchers, clay jugs, mortars, tripod chairs, sewing machines, carpets, weaved clothes, and many other ethnological items. They were made of textile, wood, ceramics, metal, copper, glass.

The items were obtained through research, as a gift or purchased from abandoned houses in rural settlements. They now belong to the MI City Museum of Negotino. The museum ethnological items are valorized as a general collection of ethnological items and some of them are featured in the ethnological exhibition which draws attention from the general public especially from the younger population.

The wine making tradition which originates and has been known to exist in the region of the Municipality of Negotino since ancient times, is concentrated in the premises of the ethnological exhibition in the City Museum of Negotino. The museum as an institution in the field of culture, presents and transmits i.e. interprets the wine making tradition to the population that visits the museum in order to get acquainted with this tradition and the cultural heritage of this municipality. A holiday celebrated by the city and connected to the wine making tradition is the day of St. Trifun, which is considered to be the patron saint of the city and is celebrated every year on 14th February, when the pruning back of the grape vine starts. At the museum you can see a lot of items representing this tradition which are part of the customs and rituals. Since Negotino is a famous wine making region, every household and especially the wineries celebrate this holiday paying homage to the patron saint.