A cryosurgical instrument and a method for separating a tissue sample from surrounding tissue of a biological tissue that is to be treated. The cryosurgical instrument includes a probe for guiding a probe head on to a biological tissue to be treated, and gas conduits for delivering coolant gas. The probe head is designed so that, in order to collect a tissue sample, a limited region of the tissue is cooled by means of the gas delivered and is separated from the surrounding tissue in a state in which it is frozen on the probe head. The instrument includes a support tube or hose 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 counter-supported during separation of the tissue sample.