The premium Boots Opticians - created in a £600,000 refurbishment - has transformed the empty former Paperchase store at the corner of Chapel Walk.
The modern eyewear boutique stocks luxury brands such as Mulberry, Armani, Hugo Boss and Kate Spade in a dedicated showroom.
But the retailer has also retained the renowned customer service it has developed in over 14 years of operating on the top floor of the historic Boots store just metres away.
Franchise owner David Blenkinsop said: “We have thousands of customers who travel to us from across South Yorkshire and Derbyshire for their eye care.
“They come back because of our attentive service and knowledgeable staff.
“At our new custom-made store, we offer so much more choice when it comes to glasses’ styles and frames, and extra space to help even more people.”
The Boots Opticians team has a clinical team of ten, including expert optometrists.
A grand store opening will take place this Friday, April 25, with Sheffield fundraising legend John Burkhill cutting the ribbon.
The store is the first new one to open on Fargate since its major revamp to install new paving, landscaping, planting, seating and lighting was completed.
Work on a new £7m events hub is due to start later this year, and a new pub is expected to open in the former Yorkshire Bank building this summer.
David added: “After 14 years here we are excited to be staying, growing, and investing in Fargate.
“Our new store keeps us in the heart of everything and is a major vote of confidence in the street.”
The new glass-fronted boutique has private test rooms, a contact lens room, glasses showroom and an audiologist service.
It has been funded in part by a £35,000 ReNew grant from Sheffield Council, which helps businesses renovate empty shops and underused spaces.
Coun Ben Miskell, Chair of the Transport, Regeneration and Climate Policy Committee at Sheffield City Council, said: “I’m thrilled to be able to welcome Boots Opticians to Fargate signalling yet another vote of confidence in the transformation we have brought to the popular street.
“With that regeneration work coming to an end, Fargate will be able to continue being the busy, thriving place it has always been.
“Sheffield city centre is an amazing place, filled with shops, cafes, restaurants and entertainment venues.
“Large parts, including Fargate, Castlegate and our Heart of the City development are adding to the appeal and attracting people to live, work and enjoy every day.
Businesses like Boots Opticians can only expand on that appeal and help Sheffield continue to thrive for many years to come.”
The existing Boots store on High Street, which dates back to 1898, will continue to serve customers their health and beauty needs.