A tissue snap-freezing apparatus without sacrificial cryogens

Molecular technologies in cancer diagnosis require a fresh and frozen tissue, which is obtained by means of snap-freezing. Currently, coolants such as solid carbon dioxide and liquid nitrogen are used to preserve good morphology of the tissue. Using these coolants, snap freezing of tissues for diagnostic and research purposes is often time consuming, laborious, even hazardous and not user friendly. For that reason snap-freezing is not routinely applied at the location of biopsy acquisition. Furthermore, the influence of optimal cooling rate and cold sink temperature on the viability of the cells is not well known. In this paper, a snap-freezing apparatus powered by a small cryocooler is presented that will allow bio-medical research of tissue freezing methods and is safe to use in a hospital. To benchmark this apparatus, cooldown of a standard aluminum cryo-vial in liquid nitrogen is measured and the cooling rate is about -25 K/s between 295 K and 120 K. Sufficient cooling rate is obtained by a forced convective helium gas flow through a gap formed between the cryo-vial and a cold surface and is therefore chosen as the preferred cooling method. A conceptual design of the snap-apparatus with forced flow is discussed in this paper.