Around a dozen members of staff formed a line on each side of the aisle to say goodbye to their loyal clientele, many of who have shopped at the Kirkcaldy shop for decades.
And there were tears shed as the doors were locked and staff embraced each other.
Among the final customers was Jessie Cairney (81) who was born just a few months before M&S opened in the High Street and has been a regular customer all her life.
She said: “I lived in Tolbooth Street when I was a girl and I was in the shop almost every day of my life.
“I feel very sad that it has closed.”
Celia Proctor and friend Dave Reid enjoyed a last hot chocolate in the cafe.
She said: “I felt really sad walking out the door.
“Marks & Spencer was the thing for me which held the High Street together.
“I’ve shopped here since I was 15 and this is a sad day.”
The very last customer to leave was Doreen Dallas (69) who lives in Kirkcaldy.
“I feel very sad,” she said.
“Not just for the customers but the staff who were exceptionally nice.
“There’s nowhere decent left to buy nice food in the town centre any more.
“I have been coming here for years, since Tesco closed because it was the only place to get food.”