The invention relates to a cryosurgical instrument and to a method for separating a tissue sample from surrounding tissue of a biological tissue that is to be treated. The cryosurgical instrument comprises a probe for guiding a probe head onto a biological tissue to be treated, and gas conduits for delivering coolant gas from a gas source of a cryosurgical apparatus to the probe head and for removing the coolant gas from the latter, wherein the probe head is designed in such a way that, in order to collect a tissue sample, a delimited area of tissue can be cooled by means of the delivered gas and can be separated from surrounding tissue in a state in which it is frozen onto the probe head. The instrument is intended to permit reliable removal of a tissue sample without damaging the tissue and to ensure a high degree of safety for the patient. To this end, the instrument has a support means in which the probe is guided and which can be moved relative to the probe in such a way that the surrounding tissue can be supported by the support means during separation of the tissue sample. The corresponding method carried out by means of the cryosurgical instrument involves separation of a tissue sample from surrounding tissue of a biological tissue that is to be treated.