A new report says one in six people have experienced negative comments or behaviour from colleagues after coming out at work.
Stonewall Scotland say employers have shown a commitment in recent years to being more inclusive.
But they point to new research that highlights continuing problems for LGBT staff.
A survey of almost 800 LGBT employees found that one in six have been discriminated against by colleagues.
Two in five trans employees have experienced the same type of behaviour from customers or clients.
The charity wants employers to adopt a zero tolerance approach and makes sure it is clear how to report anti-LGBT bullying.
The benefit, they say, will be happy staff who perform better.
Stonewall Scotland director Colin Macfarlane said: “Over the last few years employers across Scotland have shown real commitment to their LGBT staff by taking action to become more LGBT-inclusive workplaces.
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