•Quantitatively study charring progress with multi-indicators;•Analyzing the microstructure changes of wood/charcoal by measuring the thickness of tracheid cell wall and the lumen area during the charring process for the first time;•Comparing the essential indexes of experimental charred wood with archaeological charcoal fossil;•Providing key physical and chemical parameters in the wood charcoalification process. Charcoal is commonly preserved in both natural and artificial sediments, and is intensively used in paleontological, paleoenvironmental, and archaeological studies due to the abundant bio-information it contains. The biochemical properties of charcoal are also used for paleoclimatic reconstruction; however, the reliability of this approach has been challenged due to a lack of clarity on how physicochemical properties change during the charring process, as well as the temperatures required for charcoalification. To address this lack, in this study, Qinghai spruce and Chinese pine wood samples from the northeastern Tibetan Plateau were heated at different temperatures and for different lengths of time under restricted oxygen conditions. The reflectance; carbon, nitrogen, and oxygen content; and tracheid morphology were quantified before and after heating to assess changes related to the charring process. Archaeological charcoal remains were then evaluated to determine the charcoalification temperatures by comparing with the experimental results. The minimum temperature required for wood charcoalification was ∼300°C, while temperatures recorded by archaeological charcoal were concentrated at 400–500°C. During the charring experiments, the tracheid cell walls gradually homogenized, and tracheid cell wall thickness and lumen area decreased by ∼20%. On average, 50% mass losses were observed; the carbon and oxygen content (% wt.) approximately changed from 47% to 60% and 48% to 35% respectively, while the nitrogen content (% wt.) fluctuated around 0.2%. The reflectance increased slightly from 0% to 0.5%. We propose that the charcoalification of wood tissue refers to charring (in restricted air) and carbonization (in the almost absence of air) when the wood is exposed to a heat source, which then finally transforms into a black, inert solid. This quantitative study provided valuable data and a thorough assessment of the process of wood charcoalification, as well as accurately estimated the feasibility of using charcoal physicochemical properties in paleoclimatic research. Display omitted