Assisi Day Trip en route from Florence to Rome

Not all that practical (at least 5 hour train time, not much less if driving), but it can be done if it's important to you. There are direct trains to Assisi, and from Assisi to Rome.

I wouldn't bother with a guide, but that's just the way I travel. Assisi is a pretty place, especially in the evening.

No personal experience with luggage storage. This seems to be an option:
https://radicalstorage.com/

We did a Road Scholar tour of Tuscany and Umbria last year that was special, because we visited smaller cities and towns in those areas, including Assisi.
Assisi was one of our favorite places, but it seems to me that it would be quite a detour from Florence to Rome.
Orvieto would be more geographically in line with such a trip. Orvieto is great, but I know it is not Assisi.
One more thing, is that we spent more time in Assisi than Orvieto. Not sure how many hours you plan to tour Assisi?

I much preferred Orvieto to Assisi. The Orvieto Cathedral is stunning.

I spent a night in Assisi and was able to leave my large bag at a shop in the train station. The station did not have a left luggage facility. This was 10 years ago so if you decide to visit, be sure to get up-to-date info. It's about a 3 hour train ride to Assisi. You'll probably be near the Florence train station but when you get to Assisi, after a few minutes to drop your luggage, you'll need to take the shuttle bus or a taxi up to the town. After your visit, it's another bus or taxi down to the train and another 3 hours (give or take) to Rome and a taxi to your hotel. Depending on the timing, you may have a train change on one or both journeys.

The current midweek train schedules have a train that leaves Florence at 6.35 am and gets to Assisi 3 hours later with one change. The next train is at 8 am, arrives 10.40 am. So you either get up really early or you start sightseeing around 11 am. There's a train that leaves Assisi at 6.12 pm, the next isn't until 10 pm. You'd have to leave the town center around 5.30. You can buy food at the station in Florence and eat breakfast on the way (surely better than what you can buy on the train). Lunch? grab something quick in Assisi. Dinner? I guess you'll have to buy something in Assisi to eat on the train. Or will you want to try to get something in Rome at 9.30-10 pm after checking into your hotel?

For me this is an easy decision. Leave Assisi for another trip.

Driving is tough too. You will be on the road for 4-5 hours just traveling. Maybe skip Amalfi and go to Assisi (Umbria is amazing, Spello, Gubbio, Perugia). There are several towns with St. Francis history there.

Assisi is easy for a self-guide, especially with the RS book & audio tour. We listened to the story of Giotto while in the upper Cathedral.

If you make it to Assisi, don't miss Saint Carlo Acutis in Santa Maria Maggiore. He was just canonized a few months ago.

Well, I can tell you that I went to Assisi on a day trip from Florence about 25 years ago. My friend and I took the train, leaving early in the morning, and wound up arriving at Assisi about three hours later.

We had a wonderful day there and while it was a bit tedious spending three hours on the train each way, we didn’t regret it a bit. I loved Assisi and thought it was really lovely. And I wouldn’t think you would need a tour. We made our way around just using a guidebook and we’re fine. It’s a really beautiful place.

Sometimes you don’t mind putting up with some inconveniences if you’re really set on seeing something. 😊

You need more than three hours in Assisi.

That would be too cumbersome for my taste. Maybe if you were traveling by car, but by train I’m not sure. I’ve driven from Florence to Assisi many times, and it takes a bit over 2 hours, but I lived close to the Firenze Sud freeway entrance. By train it’s 3 hours and the Assisi station is down the valley at S.Maria degli Angeli, from where you need to take a taxi (or a bus). The issue in your case is also the luggage, that you would need to store somewhere and I don’t know if there are shops providing the service at S. Maria. All in all it would require a very early start from Florence, catching the 6am train (so a 4 something am wake up) for a couple hours visit at best, if you want to make it to Rome at a decent time. If I were determined to see Assisi at all costs I would borrow a night from another destination you are going.

You need more than three hours in Assisi.

Even on a day trip you could get more than three hours in Assisi. When I went, we left early in the morning and got there around 10:30 a.m. We spent the rest of the morning and most of the afternoon there. I think we left around 5:00 or 5:30 p.m. so we had plenty of time to see the town. Obviously if you spent the night there, you would have more time and that would be a good thing. But I'm just responding to the OP and saying that it is feasible, although it would require some maneuvering and some time spent on the train. But in my opinion it's worth it to see Assisi, especially if they don't think they will be back.

Great minds! We will be in some of those areas the same time. We will be in Assisi for 4 nights. We have never been before, and would not be staying this long, but we are going for the Calendimaggio. I would think even a few hours of this experience would be great. Also, chances are the crowds will be where the activities are, maybe leaving other areas less crowded. I know the hills will make traveling around slower in general. Unless maybe powerwalking uphill is your thing :). https://www.italyscapes.com/events/umbria/assisi/traditional-festivals/calendimaggio-2026-complete-guide/