Reports emerged this week of a dressing room bust-up between captain Abdoulaye Faye and midfielder Glenn Whelan at half-time and full-time of Sunday's 7-0 defeat at Chelsea.
That followed a leaked story in December which alleged Pulis and striker James Beattie had a physical altercation at Arsenal during a row over the players' Christmas party.
Meanwhile Stoke's last two matches have seen forwards Tuncay Sanli and Dave Kitson, who also swore at Pulis, march straight down the tunnel after being substituted.
Pulis was furious about the latest leak and, although he has yet to prove the identity of the source, he insists such behaviour has no place at the club.
"We have no concrete evidence who it is but we are desperately disappointed because the dressing room is a good dressing room," said the Potters boss.
"Securing Premier League status for the second year with seven games to go shows how strong the spirit has been.
"You don't do that if you have bad spirit in the dressing room and people are not pulling together.
"It is one or two precious people who have had things not gone their way who have decided they want to do certain things - it is up to them but it has no place in our dressing room.
"Glenn and Aby are really disappointed the story came out. Once it was over it was over and you just move on.
"The players have been desperately disappointed. There are some good characters in the dressing room and I believe one or two will have sorted that out.
"Everything that happens in the dressing room should stay in the dressing room and unfortunately it hasn't.
"Ever since I started professional football at 15 there was always that togetherness and solidarity in the dressing room - it is a sanctuary.
"When I started football everyone believed it."
Pulis, who held a meeting with his players on Monday, was also annoyed by the reaction of Kitson after his substitution at Stamford Bridge but excused Tuncay's petulance the previous week at home to Bolton.
"We've had a good meeting on Monday and it has been dealt with," he added.
"Tunny is an exception in some respects because he comes from a different culture and has a different way of being brought up.
"For British players to act like that, they know what the score is and if anyone has a gripe I've got a manager's room and they can knock on my door and have a chat.
"That is what we used to do when we were players; none of us were happy being brought off but we never threw our hands up in the air and tried to curry favour with supporters. We used to get on with it and go and see the manager on a Monday face-to-face.
"I've always been a bit old fashioned and thought the best way to sort things out man-to-man.
"For me it is petulant, precious people. They have to realise it is not about them, it is about the club.
"They don't let themselves down, they let the team down, the supporters down and the football club down."