Barbara Mazzolai was born in Livorno and obtained her degree with honours in Biology from the University of Pisa in 1995. From 1994 to 1998 she worked at the Institute of Biophysics of the National Research Council (CNR) where she was devoted to environmental issues and particularly to mercury pollution.  Barbara continued her studies with a masters course in Environmental Management and Control (EMAS) at the Scuola Superiore Sant’Anna in Pisa (SSSA). She then became an assistant researcher at the Centre for Research in Microengineering (CRIM Lab) from 1999 to 2004. At the CRIM she founded a new field of research on “microtechnologies and robotics for application in the environment and food industry”. From 2004 to 2009 she worked as assistant lecturer of Biomedical Engineering at the SSSA.  In November 2009, Barbara became team leader at the Centre of Micro Bio-Robotics of the Italian Institute of Technology (IIT) of Genoa, based in Pontedera, Italy, and in 2011 she was appointed coordinator of the centre. In 2011 she received her PhD in Microsystems Engineering at the Tor Vergata University of Rome.

Barbara has since been a protagonist in many frontier studies, first and foremost the study of “Dustbots”, i.e. street cleaning robots, when she was a researcher at the SSSA. While DustClean cleans and disinfects the streets, Dustcart is a small green robot that, after receiving a phone call, arrives in front of house doors and collects waste bags to put them in the wheelie bins. In 2010 Barbara received the “Marisa Bellisario” award for her scientific work and her management of the  “Dustbot” project to improve urban hygiene and the citizens’ quality of life..

Another field where Barbara has gained a name is the construction of plant or animal inspired robots. Her first project led to the creation of “Octopus”, a robotic arm that works in water by imitating the arm of that animal. In 2012 she began another project as scientific director of the first example of a futuristic project financed by the European Commission. “Plantoid” is inspired by plants and aims to create plantoid robots that are capable of imitating the behaviour of roots for exploring the earth and for environmental monitoring.

In 2013 Barbara received the Medal of the Senate of the Italian Republic and in 2015 the international prize Genova, as a woman of excellence for her research in the field of bio-inspired robotics. Also in 2015, she was nominated by RoboHub to be among the 25 most influential women in the robotics sector. From January to July 2017 she was Visiting Faculty at the Aerial Robotics Lab of the Department of Aeronautics, Imperial College London.

Barbara Mazzolai is one of the 100 female researchers invited to be part of the "100 Women for Science" project.