Cristiano Ronaldo and Co. will be looking to gain revenge for their loss to Diego Simeone's gritty side in the Copa del Rey final in May, while both teams are hoping to get ahead in the frantic race at the top of the La Liga table.
The outspoken Simeone recently proclaimed Real Madrid and Barcelona to be on a completely different plane to every other team in La Liga.
"Madrid and Barcelona play in a different league," Simeone said at an August press conference, as reported by Sky Sports.
"This is a boring championship for the rest of the supporters who can only aspire to be third or fourth," he said.
Such comments would seemingly indicate that Simeone gives Atletico little chance of toppling their archrivals on Saturday night, but few in the Real Madrid camp will consider themselves overwhelming favorites.
Nor will anyone expect Simeone to approach the task with anything other than total commitment. The Argentine will lock horns with new Real manager Carlo Ancelotti, and both will be looking to exploit perceived flaws in their opponent's tactical setup.
This will be a battle not just of individual players, but between two coaches searching for the perfect balance in their starting 11s.
Here are what we expect to be some of the key matchups of an enthralling derby.
He may only be a newcomer to the Real Madrid setup, but 21-year-old Isco has already established himself as a key attacking weapon in the side.
The former Malaga man has started every game under Ancelotti, playing in an advanced midfield role behind striker Karim Benzema.
Isco provides a huge threat himself, as evidenced by his tally of four goals in six matches, but also operates as the hub, linking play between Real's other danger men, Benzema, Cristiano Ronaldo and Gareth Bale or Angel Di Maria.
Shutting down Isco, and thus taking control of the area of the pitch which he patrols, will be a critical task for Atletico if they are to take some of the sting out of their opponent's attacking force.
Simeone will understand very well that his players must press with intensity all over the pitch, but Mario Suarez and Gabi in particular will have to be at their energetic best.
As Isco buzzes around, the two central midfielders will likely take it in turns to stick with him, with Suarez likely to stay deeper and Gabi to move further up the pitch when required. Koke as well may be asked to help out, though he is likely to be toiling to contain the dangers down the left-hand side.
Atletico may have lost their magnificent Colombian striker Radamel Falcao to Monaco, but thus far David Villa and Diego Costa have done a sterling job up top for Simeone.
Costa, in particular, is in sensational form, having cracked seven goals in six matches in the league.
The Brazilian was recently selected as having the second-biggest ego in La Liga by B/R's Kieran Sobels (you can probably guess who came first). Costa loves a scrap. Probably a little too much, in fact, as he quite often finds himself in strife, as Sobels points out.
Sergio Ramos, the man most likely to be marking Costa, is no shrinking violet himself. He also happens to be a defender right out of the top draw, so this battle of egos, of temperaments and of footballing abilities is likely to be a fascinating one.
There is a long history of niggle between the two, via Mail Online, and watching the next chapter of their rivalry unfold could be one of the most entertaining elements of the match.
How does one stop Cristiano Ronaldo?
The fact that the prancing Portuguese has knocked in 208 goals in 205 games for Los Blancos suggests nobody has yet found a suitable answer to that question.
The former Ballon d'Or winner is undoubtedly going to be running at Atletico from all angles, but the Rojiblanco right-back Juanfran will likely be the man attempting to stifle his progress down Real Madrid's left edge.
The Spanish international has had a patchy start to the season and looked particularly vulnerable against Zenit St. Petersburg in the Champions League.
He will require assistance from Koke and Mario Suarez, in containing Cristiano, but even then the former Manchester United player could prove too much to handle.
Juanfran will need to be at his absolute best to prevent Ronaldo running amok.
There was speculation that gangly winger Angel Di Maria might leave Real Madrid, via Gary Jones of The Daily Star, in the summer to facilitate Gareth Bale's arrival, but the Argentine opted to remain at the club to fight for a first-team place.
So far that decision has been justified by several excellent performances from the Rosario Central product.
Ben Hayward, writing for Goal.com, underlines how decisive "Noodle" was against Getafe last weekend:
Di Maria tracked back on countless occasions, worked tirelessly to recover the ball and was an omnipresent force as Madrid poured forward.
While Benzema was booed, the winger won the applause of the home fans for his unquestionable commitment and proved a constant threat to the visiting defence.
The Argentine also helped set up the first goal and was unfortunate not to score one of his own as he hit two venomous efforts just off target. Such was his work-rate, in fact, that Marca later described the winger as "the soul" of the Madrid side.
Bale, meanwhile, if handed a start, will be especially keen to show how influential he can be, in the wake of stiff competition from his own teammates.
Atletico left-back Felipe Luis must ensure that whichever of these two big-eared fliers he comes up against, he does not allow him the time or space to gather speed and momentum.
Either man has the ability to win the game for Real Madrid if he gets the wind in his sails.