Reduced thiol compounds (RTCs), such as glutathione (GSH), cysteine (Cys), and homocysteine (Hcy), are important components of many amino acids and proteins. They are also basic building blocks that make up organisms and play an important role in physiology and pathology. Therefore, based on the fundamental characteristics of RTCs in vivo, researchers have reported their use in biosensing and imaging analysis and as carriers in life sciences and health sciences. In this review, we will introduce the principles of RTCs and highlight their applications in biosensing and imaging analysis and as carriers reported from 2015 to 2020. Yingshu Guo (top left) obtained her Ph.D. degree from Qingdao University of Science and Technology in 2012. She is currently a professor at Linyi University, China, with research interests in bioanalytical chemistry and chemical biology. Xiaofei Zheng (top middle) obtained his B.S. degree from Linyi University in 2019. Then he pursued his M.S. degree under the supervision of Prof. Yingshu Guo at Linyi University. His current research is focused on bioanalytical chemistry. Xiuping Cao (top right) obtained her B.S. degree from Shandong University of Traditional Chinese Medicine in 2019. Then she pursued her M.S. degree under the supervision of Prof. Yingshu Guo at Linyi University. Her current research is focused on bioanalytical chemistry and chemical biology. Wenxin Li (bottom left) obtained her B.S. degree from Qilu Institute of Technology in 2020. Then she pursued her M.S. degree under the supervision of Prof. Yingshu Guo at Linyi University. Her current research is focused on bioanalytical chemistry and chemical biology. Di Wu (bottom middle) obtained her B.S. degree from Linyi University in 2020. Then she pursued her M.S. degree under the supervision of Prof. Yingshu Guo at Linyi University. Her current research is focused on bioanalytical chemistry. Shusheng Zhang (bottom right) obtained his Ph.D. degree from Nanjing University in 1999. He is a leading researcher at Linyi University with research interests in bioanalytical chemistry.