Taking the bus is the cheapest way to travel from Florence to Bologna. Most buses traveling from Florence to Bologna depart from either Florence's Piazzale Montelungo or Autostazione Busitalia-Sita Nord and take an hour and a half to arrive at Bologna's central bus station. Most buses depart from Montelungo, which is a short walk to the north of the historic center of Florence. At least 10 buses travel from Florence to Bologna every day, with the earliest departing at 2:10 a.m. Bus companies that travel from Florence to Bologna include Flixbus, Baltour, and BusCenter. Ridesharing services like BlaBlaCar also frequently travel from Florence to Bologna.

Since there are no direct flights, the fastest way to travel from Florence to Bologna is by taking a high-speed train. Trains traveling from Florence to Bologna depart from either Florence's Santa Maria Novella, Rifredi, or Campo di Marte train stations and take as little as 35 minutes to arrive at Bologna Centrale train station. Both Trenitalia and Italo offer frequent trains from Florence to Bologna, with the first train departing at 4:32 a.m. Nearly 90 trains travel from Florence to Bologna per day.

Taking the train is the most popular way to travel from Florence to Bologna. Taking a high-speed train is usually an hour faster than taking the bus, while also only costing an additional €6 on average. Be sure to look for a Frecciarossa, Frecciargento or Italo train, which are approximately an hour faster than taking a regional Trenitalia train. Trains traveling from Florence to Bologna are also usually more convenient for travelers since approximately 90 trains depart from Florence throughout the day.