Introduction. The contact point between an astrocyte and a chemical synapse is the location of the signaling and transport processes that play an important role in functioning of the nervous system and in neurological diseases pathogenesis of humans and animals. The goal of the study was to investigate the correlation between the average size of the active zone of the synapse and the frequency of synapse-astrocyte contact formation in the cerebral cortex layers of rats.Materials and methods. The 40 μm frontal sections of the primary somatosensory cortex of 5 outbred male white rats were taken as the material for the study. Astrocytes were labelled for Transmission Electron Microscopy (TEM) by incubation of the tissue sections with primary antibodies to the s100β protein and with horseradish peroxide-conjugated secondary antibodies, followed by the development of the label in the reaction with DAB. For each layer, 250 images were obtained at 25,000х magnification. On the images the length of the synaptic cleft was measured and the number of synapses forming contact with the astrocyte membrane was counted.Results. The value of the fraction of the chemical synapses forming contact with the astrocyte membrane in the primary somatosensory cerebral cortex of rats demonstrated the direct correlation with the average length of the synaptic cleft only in the first to fourth layers. Thus, in the first layer, the value of the fraction of synapses forming contact with the astrocyte was the smallest (PI = 0.27 ± 0.1), as was the length of the synaptic cleft (lI = 329.45 ± 10.45). When moving deeper into the cortex, the fraction of synapses forming contact with the astrocyte and the length of the synaptic cleft increased from the second (PII = 0.48 ± 0.11 and lII = 363.64 ± 11.14) to the third layer (PIII = 0.69 ± 0.09 and lIII = 382.27 ± 9.81), followed by the decrease of both values in the fourth layer (PIV = 0.53 ± 0.09 and lIV = 355.2 ± 8.12). In the fifth layer, the fraction of synapses forming contact with the astrocyte sharply increased again (PV = 0.68 ± 0.08), which, however, was not accompanied by the proportional increase of the average length of the synaptic cleft (lV = 350.79 ± 7.82). At the same time, in the sixth layer of the cortex, on the contrary, the sharp increase in the average length of the synaptic cleft (lVI = 396.03 ± 10.77) was observed, reaching the maximum value through the cortex, with low, compared to other layers, fraction of synapses forming contact with the astrocyte (PVI = 0.44 ± 0.09). Thus, it turned out that the fraction of synapses forming contact with the astrocytic membrane is more related to the density of the astrocytic membranes in the layer (the research we published earlier), rather than to the average length of the synaptic cleft. Discussion and conclusions. The obtained results substantiate that the contact is formed as a result of a combination of random membranes encounter followed by the selective anchoring or repulsion of the astrocyte membrane under the influence of various factors, only partly determined by the size of the chemical synapse.