British Transport Police will be patrolling stations and trains when the night Tube launches next weekend.
Transport for London (TfL) said the deployment of officers will "help customers travel with confidence" as the all-night service begins on the Central and Victoria lines on Friday and Saturday.
The deployment is part of an investment by TfL of an extra £3.4 million towards policing for the night Tube.
It will see around 100 officers out on the network when full services are under way, with at least as many officers out during the night as would be seen during the day, TfL said.
A BTP control centre is co-located with London Underground's control centre, both of which operate 24-hours a day.
Officers will be supported by BTP rapid-response vehicles spread across London and by more than 12,000 CCTV cameras across the network.
The Metropolitan Police, responsible for policing beyond the Tube stations, will also be at key hubs and bus stations.
The first two Tube lines, Victoria and Central, start operating night Tube services at 51 stations on Friday and Saturday nights from next weekend.
Mayor of London Sadiq Khan said: "The opening of the night Tube marks the start of an exciting new chapter in London's life. It will unlock the full potential of London's night time economy and support the thousands of Londoners who travel to or from work at night.
"Keeping Londoners safe is my number one priority. The extra investment in British Transport Police officers will ensure that all Londoners using the night Tube feel safe, whether getting back from work or going out at the weekend with friends.
"Passengers on the night Tube must be able to travel with the same confidence they are used to during the day. That's why we're investing £3.4 million to ensure that dedicated officers are on hand to offer the support and visible reassurance Londoners expect."