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The Maidstone and Tunbridge Wells NHS has replaced a traditional black and white zebra crossing at its hospital in Barming with a rainbow coloured one.
Trust chief executive Miles Scott said: "It's an demonstration that we are open to everybody - we want all of our staff and all of or patients to feel welcome here."
KMTV's Kristina Curtis reports on the new pedestrian crossing
The rainbow is a symbol of the LGBT (lesbian, gay, bisexual, and transgender) community.
Mr Scott said: "We've had a terrific response. Many people understood the symbolism. Others have asked us the purpose, which has given us the chance to explain it to them.
"We can't expect our staff to give great care to our patients if they don't themselves feel welcome and happy here."
Alex Beardmore is a member of the LGBT community and a radiographer at the hospital.
He said: "It's brilliant. This is the first Rainbow crossing in Kent and the first for the NHS.
"It's so vibrant and also it's so much safer. You can't miss it!"
Mick Stupples, co-chairman of the Maidstone and Tunbridge Wells NHS Trust’s LGBT Network, said: “The trust’s LGBT staff network has been working hard to increase awareness of LGBT issues and improve the experiences of healthcare for LGBT staff and patients.
“While we’ve come a long way as a society in recent years to be more inclusive, there’s still more we can do to.
"The Rainbow crossing acts as a visual symbol of inclusion and enhances the work we’ve been doing to better support our LGBT staff and patients.”