BACKGROUND With the increasing global incidence of diabetes mellitus (DM), diabetic retinopathy (DR) has become one of the leading causes of blindness in developed countries. DR leads to changes in retinal neurons and microcirculation. Rtx1TM (Imagine Eyes, Orsay, France) is a retinal camera that allows histological visualisations of cones and retinal microcirculation throughout the DM duration. OBJECTIVE This study aimed to analyse the cones and retinal microvascular changes in 50 diabetic individuals and 18 healthy volunteers. The patients participated in the initial visit and two follow-up appointments, one and two years after the study, beginning with Rtx1TM image acquisition, visual acuity assessment, macular OCT scans and blood measurements. RESULTS The study revealed significant differences in the cone density, mosaic arrangement and vascular morphology between healthy and diabetic patients. The final measurements showed decreased photoreceptor and microvascular parameters in the DR group compared with the control group. Furthermore, in the 2-year follow-up, both groups' Rtx1TM-acquired morphological changes were statistically significant. CONCLUSIONS Rtx1TM technology was successfully used as a non-invasive method of photoreceptors and retinal vasculature assessment over time in patients with diabetic retinopathy. The study revealed a trend toward more vascular morphological changes occurring over time in diabetic patients.