PURPOSE: The primary healthcare setting is well placed for health screening. Tear fluid composition gives valuable information about the eye and systemic health, and there is now significant interest in the potential application of tears as a tool for health screening; however, the acceptability of tear collection in the primary healthcare setting as compared with other methods of human sample collection has not been previously addressed. The objective of this study was to evaluate the patient acceptability of tear collection in a primary healthcare setting. METHODS: This was a cross-sectional study on 383 adult patients seeking primary healthcare, who were not diabetic and were not attending for an eye-related complaint. Tear collection was done using Schirmer strips, and an interviewer-administered questionnaire was conducted to collate information on the pain score (0-10) of the Schirmer tear collection, as well as to score the pain associated with their previous experience of antecubital venous puncture and finger prick test. RESULTS: The pain score for Schirmer tear collection was significantly lower (p < 0.001) than antecubital venous puncture but higher (p < 0.001) than finger prick. The pain scores for all three procedures were significantly higher in participants of younger age, female gender, and higher education level. Among the participants, 70% did not mind their tears being collected to screen for eye problems, whereas only 38% did not mind this procedure being performed for general health screening. Nevertheless, 69% of the participants preferred tear to urine collection, and 74% of participants preferred tear to blood collection. CONCLUSIONS: Tear collection using Schirmer strips is a highly acceptable form of investigation that has the potential for use in health screening in the primary healthcare setting. This study has implications on using tear collection as a method of ocular and systemic health screening in the primary healthcare setting.