<p>The modelling of peatland functioning, in particular the impact of anthropogenic warming and direct human disturbance on CO<sub>2</sub>, CH<sub>4</sub> and N<sub>2</sub>O, requires detailed knowledge of the peat structure and of both water and gas flow with respect to the groundwater table level. To this end, freezing is nowadays increasingly used to obtain small size peat samples for X-ray micro computed tomography (X-ray μ-CT) as required by the need to increase the resolution of the 3D X-ray CT images of the peat structure recorded. The aim of this study was to analyze the structure of a peat material before and after freezing using X-ray μ-CT and to look for possible alterations in the structure by investigating looking at the air-filled porosity. A highly decomposed peat material close to water saturation was selected for study and collected between 25 and 40 cm depth. Two samples 4 × 4 × 7 cm<sup>3</sup> in volume were analyzed before and after freezing using an X-ray μ-CT Nanotom 180NF (GE Phoenix X-ray, Wunstorf, Germany) with a 180 kV nanofocus X-ray tube and a digital detector array (2304 × 1152 pixels Hamamatsu detector). Results showed that the continuity and cross section of the air-filled tubular pores several hundreds to about one thousand micrometers in diameter were altered after freezing. Many much smaller air-filled pores not detected before freezing were also recorded after freezing with 470 and 474 pores higher than one voxel in volume (60 × 60 × 60 μm<sup>3</sup> in volume each) before freezing, and 4792 and 4371 air-filled pores higher than one voxel in volume after freezing for the two samples studied. Detailed analysis showed that this increase resulted from a difference in the whole range of pore size studied and particularly from a dramatic increase in the number of air-filled pores ranging between 1 voxel (216 10<sup>3</sup> μm<sup>3</sup>) and 50 voxels (10.8 10<sup>6</sup> μm<sup>3</sup>) in volume. Theoretical calculation of the consequences of the increase in the specific volume of water by 8.7 % when it turns from liquid to solid because of freezing led to the creation of a pore volume in the organic matrix which remains saturated by water when returning to room temperature and consequently to the desaturation of the largest pores of the organic matrix as well as the finest tubular pores which were water-filled before freezing. These new air-filled pores are those measured after freezing using X-ray μ-CT and their volume is consistent with the one calculated theoretically. They correspond to small air-filled ovoid pores several voxels in volume to several dozen voxels in volume and to discontinuous air-filled fine tubular pores which were both detected after freezing. Finally, the increase in the specific volume of water because of freezing appears also be also responsible for the alteration of the already air-filled tubular pores before freezing as shown by the 3D binary images and the pore volume distribution.</p>