Background: The use of minimally invasive biomarkers (MIBsphysiological biomarkers obtained from minimally invasive sample types) has expanded rapidly in science and medicine over the past several decades. The MIB approach is a methodological strength in the field of human and non-human primate evolutionary biology (HEB). Among humans and our closest relatives, MIBs provide unique opportunities to document phenotypic variation and to operationalize evolutionary hypotheses.Aims: This paper overviews the use of MIBs in HEB. Our objectives are to (1) highlight key research topics which successfully implement MIBs,(2) identify promising yet under-investigated areas of MIB application, and(3) discuss current challenges in MIB research, with suggestions for advancing the field.