Acne diagnosis, severity assessment and treatment follow‐up rely primarily on clinical examination. In vivo reflectance confocal microscopy (RCM) provides non‐invasively, real‐time images of skin lesions with a level of detail close to histopathology. This systematic literature review aims to provide an overview of RCM utility in acne and a summary of specific features with clinical application that may increase objectivity in evaluating this condition. We used the Preferred Reporting Items for Systematic Reviews and Meta‐Analyses (PRISMA) guidelines for reporting our results. We systematically searched three databases: PubMed, Clarivate and Google Scholar (January 2022). All included studies used RCM to investigate acne in human patients and reported the investigated skin area and type (acne lesions or clinically uninvolved skin), the substance used in the case of treatment. Our search identified 2184 records in the three databases investigated. After duplicate removal, 1608 records were screened, 35 were selected for full‐text assessment, and 14 were included in this review. We used the QUADAS‐2 tool to evaluate the risk of bias and applicability concerns. RCM was selected as the index test and clinical examination as the reference standard. The total number of patients from all studies was 291, with 216 acne patients and 60 healthy participants aged between 13 and 45 years. The 14 considered studies analysed 456 follicles from healthy participants, 1445 follicles from uninvolved skin in acne patients and 1472 acne lesions. Consistent RCM findings concerning follicles of acne patients reported across studies were increased follicular infundibulum size, thick, bright border, intrafollicular content and inflammation. Our analysis indicates that RCM is a promising tool for acne evaluation. Nevertheless, standardization, a unified terminology, consistent research methods and unitary reporting of RCM findings are necessary. PROSPERO registration number CRD42021266547.