Description: Frozen tissue samples represent the state of the art in tissue preservation in many molecular analysis techniques as well as in in membrane analysis using free-fracture techniques. Rapid freezing eliminates the artifact caused by ice crystal formation within the tissues. Ice crystal nucleation and growth occurs between 0?C and -20?C typically. To avoid this damage and minimize destruction of proteins, RNA and DNA by lytic enzymes, cells or tissues have to be rapidly cooled through this temperature band. This is typically done in an isopentane bath cooled by liquid nitrogen (LN2) to -150?C. This 2 step process eliminates artifacts caused by ice nucleation as well as artifacts caused by nitrogen bubbles that surround the tissue as it boils if submerged directly into LN2. While these open methods are acceptable for terrestrial laboratories, they would not be compatible with experimentation on the International Space Station. Our proposed gaseous nitrogen-based Rapid Freezer clamp would provide an alternative means to rapid cool through ice crystal nucleation and growth temperatures without exposing the crew to the spill hazards of LN2 and chilled isopentane as well as the extreme flammability of isopentane.

Benefits: The Techshot Rapid Freezer (formally known as the Snap Freezer) is being developed as an enabling device for conducting microgravity research aboard the ISS. Beginning first as an application for NASA mission programs, Techshot will have developed a unique device capable of safely rapidly freezing samples in space. Therefore, the initial targeted application of the proposed innovation is an offering of both the equipment and services associated with flight hardware and integration activities, which are highly desired by NASA-funded scientists. In 2015, Techshot signed a Space Act Agreement with NASA, and was awarded an Indefinite Delivery Indefinite Quantity (IDIQ) contract from the agency to enable the Government to lease (as needed) a wide variety of our ?professional grade? flight hardware. This new business model is expected to be very attractive to NASA because it reduces the agency?s upfront risk for flight hardware development, and more importantly, it reduces NASA?s cost of ongoing maintenance and upkeep of the flight hardware - for the life of the equipment.

The Rapid Freezer is expected to be of particular interest to the pharmaceutical and biotechnology industries, academic researchers and terrestrial national laboratories, and Techshot is committed to investing in its commercialization. Beginning with the Phase I award, the company built a technology demonstrator with internal funds, and it will continue to invest its own funds in the further maturation of the Rapid Freezer system. While typical laboratories snap freeze tissues by plunging specimens into liquid nitrogen or an isopentane bath chilled by liquid nitrogen, this may not be compatible with all experimental protocols or facility safety standards, which will provide commercial opportunities. Much like the safety concerns on the ISS, significant burn hazards exist from liquid nitrogen spills and isopentane is an extremely flammable liquid. When these conditions exist in terrestrial labs, researchers use cold blocks to freeze samples. But even actively-cooled plates in cryostats do not typically reach temperatures sufficient for effective rapid freezing. This may not freeze sensitive or thick tissues at a fast enough rate to preserve the most sensitive DNA, RNA, proteins and crystal structures.