This review article focuses on the recent advances in fluorescent probes for detecting and imaging cancer biomarkers. It highlights the design strategies of small-molecule and nano-sized fluorescent probes, their construction methods, and applications in detecting multiple biomarkers such as enzymes, reactive oxygen species (ROS), reactive sulfur species (RSS), and microenvironments. The article emphasizes the advantages of fluorescent probes, including non-invasiveness, fast response, high sensitivity, and selectivity, which make them valuable tools for early cancer diagnosis and therapeutic evaluation. The review also discusses the challenges and future directions in the development of fluorescent probes, aiming to guide researchers in advancing the field of cancer-marker fluorescent probes from bench to bedside. Key topics include the design of BODIPY, rhodamine, and tetraphenylethylene (TPE) fluorescent probes, the use of metal nanoclusters (MNCs), fluorescent nanoparticles (FNPs), and quantum dots (QDs) for cancer biomarker detection, and the application of these probes in detecting various cancer biomarkers such as enzymes, ROS/RSS, and microenvironmental factors. The article concludes by addressing the limitations and future prospects of fluorescent probes in cancer diagnosis and therapy.This review article focuses on the recent advances in fluorescent probes for detecting and imaging cancer biomarkers. It highlights the design strategies of small-molecule and nano-sized fluorescent probes, their construction methods, and applications in detecting multiple biomarkers such as enzymes, reactive oxygen species (ROS), reactive sulfur species (RSS), and microenvironments. The article emphasizes the advantages of fluorescent probes, including non-invasiveness, fast response, high sensitivity, and selectivity, which make them valuable tools for early cancer diagnosis and therapeutic evaluation. The review also discusses the challenges and future directions in the development of fluorescent probes, aiming to guide researchers in advancing the field of cancer-marker fluorescent probes from bench to bedside. Key topics include the design of BODIPY, rhodamine, and tetraphenylethylene (TPE) fluorescent probes, the use of metal nanoclusters (MNCs), fluorescent nanoparticles (FNPs), and quantum dots (QDs) for cancer biomarker detection, and the application of these probes in detecting various cancer biomarkers such as enzymes, ROS/RSS, and microenvironmental factors. The article concludes by addressing the limitations and future prospects of fluorescent probes in cancer diagnosis and therapy.