The application relates to a cryosurgical instrument for separating a tissue sample from a biological tissue that is to be treated. The cryosurgical instrument (10) comprises a probe (40), for bringing a probe head (42) to the biological tissue to be treated, and a gas delivery device for delivering coolant gas from a gas source of a cryosurgical appliance to the probe head, wherein the latter isdesigned in such a way that, by means of the delivered gas, a limited area of tissue can be cooled, in order to recover a tissue sample, and can be separated from the surrounding tissue in a state frozen onto the probe head. The instrument is designed to allow a tissue sample to be collected reliably and in a manner that is gentle on the tissue and that provides a high degree of safety for the patient. For this purpose, the instrument comprises a support tube (60) in which the probe is guided and which is movable relative to the probe in such a way that the surrounding tissue can be supportedby means of the support tube during the separation of the tissue sample. An accelerating device (110) is also provided, which makes available a predefined force for the separation procedure.