Terry and Cole have been in a race against time to recover from ankle injuries for the west London derby at Loftus Road.
But they have trained without incident for the past three days and are set for a reunion with Anton Ferdinand, who Terry was cleared in court of of racially abusing back in July.
"They've recovered, had a couple of days training and we expect them to be available for tomorrow [Saturday]," Di Matteo said.
"We're going to respect the rules and the protocol that is in place from the Premier League and set a positive standard to the millions of people watching across the world."
He added of the handshake: "We'll go ahead with it, yes."
Di Matteo revealed he had held talks with his opposite number, and former Chelsea team-mate, Mark Hughes but did not know whether QPR's players would snub Terry and Cole.
He said: "I do not know, but I hope that they will as well respect the rules and go ahead with it and we can all focus on the football."
Hughes revealed this afternoon [Friday] that they were awaiting guidance from the Premier League, who have confirmed it must go ahead.
"There has been dialogue between the Premier League, Queens Park Rangers and Chelsea in relation to tomorrow's [Saturday] match between the two clubs," read a statement released by the Premier League.
"All parties understand and acknowledge that the pre-match handshake will go ahead as part of the normal pre-match activity."
Hughes earlier hinted at his reservations of the pre-match ritual and is exasperated that it is overshadowing tomorrow's [Saturday] highly-charged west London derby.
"There's a lot of debate every time we have a game against Chelsea," Hughes said.
"I've got my own views on the handshake and I raised those at Premier League level even before this season started. I don't want to go into that.
"We'll be guided by the Premier League and will respect the handshake if we're told that's what needs to happen.
"I'm conscious of the fact that every time we play Chelsea the issue of the handshake clouds everybody's mind and the focus is taken away from a great Premier League game.
"It's not my decision to make. We're governed by the Premier League and if we're told it will go ahead, then we will do that.
"We haven't spoken to them but I imagine they'll insist the handshake goes ahead, which isn't a problem."
Ferdinand is reported to also be considering snubbing Ashley Cole after the Chelsea left-back testified as a character witness for Terry.
"We've had a discussion with the players but that's between myself and them. We don't have to broadcast that," Hughes said.
"We'll have to wait and see (if Ferdinand shakes Terry's hand). Everybody has a mind of their own and will make a personal decision.
"It's unfortunate that they're in a situation where it's in the public domain. Anton is fine, there's no problem with him."
QPR striker Bobby Zamora has indicted he would shake the hand of Terry, whom he played alongside at schoolboy level.
"I've known John Terry for years and he's a very good friend of mine," Zamora told the Sun.
"We grew up together and of course I'll shake his hand if he's playing on Saturday."
Terry remains the subject of a Football Association investigation over allegations he racially abused Ferdinand, a charge he denies, while left-back Cole was a character witness for Terry during this summer's trial.
The Premier League yesterday [Thursday] expected the pre-match handshake to go ahead as QPR and Chelsea held talks about how to approach Saturday's fixture.
Both clubs also issued warnings to their own fans, with QPR posting on their official website, www.qpr.co.uk, under the title 'Your Behaviour: A Reminder', while Chelsea posted their own statement called 'The Right Rivalry'.
Chelsea's statement on their official website, www.chelseafc.com, read: "Last January's FA Cup match at QPR and the meeting between the sides in April at Stamford Bridge were good examples of passionate and vocal rivalry with the fans of both clubs recognising that abuse and discrimination have no place in a football stadium, nor anywhere else in society.
"Chelsea Football Club wishes to see the level of respect at those games continued throughout this campaign, as it has been in our opening matches.
"Both Chelsea and QPR will work together with the police to ensure that anyone using discriminatory or inflammatory language on Saturday is identified, and that the strongest possible action is taken against them."
Chelsea fans have also been known to taunt Liverpool about Hillsborough, most notably during a minute's silence on the anniversary of the tragedy before their side's FA Cup semi-final against Tottenham last season.
The Blues are understood to have identified and taken action against some of those involved and Di Matteo said today [Friday]: "Our message is to respect the victims' families.
"That day was a tragedy for the families, who have lost their loved ones, and football in general.
"Now there's an end to the uncertainty there was over that tragedy, we kind of have to look forward and remember the victims of the tragedy.
"Our sympathy goes out to the families who lost husbands and children and so on.
"The message to our fans is to respect that and show a good example of what our fans are like."
A Chelsea spokesman added: "Chelsea Football Club's sympathy towards those that suffered as a result of the Hillsborough Disaster 23 years ago is well known and has consistently been made clear.
"We have also stated on a number of occasions our view of Chelsea supporters who engage in inappropriate and unacceptable chants, songs or statements that dishonour all those that suffered as a result of the tragic events that day."