The freezing is needed to have the surgical sample hard enough to be cut in a thin section mounted on a microscope slide and examined under the microscope. It is the freezing that makes the procedure fast enough to be done during a surgery. It is known that frozen section gives poor results because during the freezing the water inside the tissue creates ice crystals that enlarge and deform the cellular structure and the morphology of the tissue. The report given by the Pathologist is usually limited to a "benign" or "malignant" diagnosis and communicated to the surgeon operating via intercom. When operating on a previously confirmed malignancy, the main purpose of the pathologist is to inform the surgeon if the surgical margin is clear of residual cancer, or if residual cancer is present at the surgical margin. PrestoChill addresses the primary limitation of the frozen section for accurate diagnosis. Ice artifact. The maximum crystals formation phase of water is between 0 and -5°C. With conventional freezing techniques, samples stay in this range of temperature for a long time, allowing the formation of ice crystals. Cryostat freezing is obtained by cooling through air, which is not the best heat exchange medium, liquid nitrogen creates a layer of gas around the sample that insulates it from the cold liquid and slows down the freezing. Also using another type of liquid, such as Isopentane, the sample is surrounded by the medium and it is the last part of the block that freezes. All of these techniques increase the frequency and size of ice crystals that impact the final morphology. The PrestoCHILL is an advanced pre-analytical system for producing frozen section blocks in a fast, reliable and standardized way. This system allows the production of high quality frozen tissue blocks thanks to the face down cryoembedding technique and the temperature of -40°C, the best compromise between the fast freezing and the optimal temperature for cutting. The face down Cryoembedding technique permits the direct contact of the sample with the cold metal giving the fastest freezing possible and allows the user to embed tissues for frozen sections with a level of control and predictability that can surpass even paraffin embedding. The temperature of-40°C impedes the water to freeze slowly and, at the same time, gives blocks that can easily reach the optimal cutting temperature of -20°C. With PrestoCHILL, tissues freeze significantly faster than with conventional methods and the freezing time is controlled. Moreover multiple blocks can be prepared rapidly. The blocks are prepared with the entire tissue face in a precisely flat plane, requiring minimal trimming and resulting in little tissue wastage. Another important feature of this method is the precision: since tissue adheres to the freezing temperature metal, it can be oriented in the preferred position either flat or on edge. PrestoCHILL is a complete system with all the tools needed to achieve the best freezing and the best orientation of the tissue: a dedicated kit for MOHS sample freezing is also available. MOHS When operating on previously confirmed malignancy the main purpose of the pathologist is to inform the surgeon if the surgical margin is free of residual tumor. The method of processing may be done with a bread loafing technique (CCPDMA) but margin controlled surgery can be performed using a variety of tissue cutting and mounting methods. Mohs surgery, is microscopically controlled surgery used to treat common types of skin cancer. During the surgery, after each removal of tissue and while the patient waits, the tissue is examined for cancer cells. That examination informs the decision for additional tissue removal. Mohs surgery is one of the many methods of obtaining complete margin control during removal of a skin cancer using frozen section histology. Using a scalpel, the surgeon removes a thin layer of visible cancerous tissue. Some skin cancers may have extensions that aren't visible from the surface. The lab analysis, which comes next, will determine if additional surgery is required to remove these cancer margins. The surgeon then cuts the tissue into sections, color codes them with dyes and draws a map of the surgical site. In the lab, a technician freezes the submitted tissue, after placing relaxing cuts the specimen is placed on the flat surface of the PrestoCHILL with the lower surgical margins face down. The peripheral margins are flattened on the surface. Gentle pressure is applied to the center of the sample to insure that the deep and peripheral margins are in contact with the frozen surface. When using the Mohs kit a ring mold is then placed around the specimen and the MCC medium is applied before setting the chuck on top of mold followed by the heat extractor. After one minute the specimen is removed from the mold and transferred to the cryostat where sections are taken. The technician cuts very thin horizontal slices. The slices are placed on microscope slides, stained and covered. This meticulous process takes time. Use of PrestoChill allows you to rapidly prepare blocks with great precision. Using a microscope, the surgeon examines all the edges and underside of the tissue on the slides and, if any cancer cells remain, marks their location on the map. The physician then lets you know whether you need another layer of tissue removed. Back in the operating room, the surgeon injects more anesthesia if needed and removes another layer of skin, precisely where the cancer cells remain, based on the map. This entire process is repeated as many times as needed until there are no more cancer cells. PrestoChill utilizes a facedown cryoembedding technique that allows for accurate identification of the surgical margins. This can eliminate tedious and time consuming manipulations allowing the frozen section technician to prepare the final slides more quickly. MCC (Milestone Cryoembedding Compound) is used to embed the samples and adhere them to the cryostat chuck for sectioning. MCC has been specially formulated with a lower viscosity that flows easily into the cryomolds and enhances adhesion to the chuck. The inherent limitations of frozen sections can be corrected, overcome or eliminated with the use of Milestone PrestoCHILL and support products. These limitations may include: Time: Frozen sections need to be prepared and diagnosed quickly. The PrestoCHILL allows for rapid freezing. The ease of use, ability to handle multiple samples simultaneously and the flat plane that eliminates excessive facing time all allow the technician to produce a slide quickly. Drying artifacts. A great deal of artifact in frozen sections is a result of "drying artifact". Slides immediately fixed for 15-60 seconds in FineFIX utilizing the FineFIX Module will show nuclear detail that rivals a formalin fixed paraffin embedded section. Fat. Traditional approaches to section fat or breast tissue in the cryostat have usually been imperfect at best. Fat is an inaqueous material that does not freeze at normal cryostat temperatures. Use of freezing sprays are discouraged due to the fact that tissue is unfixed and aerosols may be formed while trying to super cool the fat to achieve a section. Use of liquid nitrogen to achieve very low temperatures may harden the fat sufficiently to achieve a section but at that temperature lymph nodes or tumor may shatter. PrestoCHILL allows optimized sectioning of the tissue of interest as well as fatty margins to be sectioned. Best operating practices are available as video tutorials for further instruction. The quality of the section is inferior. In the hands of a well trained and experienced technician a frozen section can be cut and stained with the quality of a permanent section. This requires optimized orientation, optimized freezing temperatures and fixation of the slides immediately upon acquiring the section. Milestone PrestoCHILL and FineFIX module can optimize and standardize freezing temperature and time parameters to achieve consistent results. Flash Freeze for Biobanking. There are few resources as valuable to cancer researchers as tissue samples. The biospecimens that patients donate to aid in the search for cancer related biomarkers. Harvesting ad storing human tissue for research purposes is not new. Human bio-specimens have been collected and stored for 100 years, and today, in excess of 300 million biospecimens are warehoused in freezers or stored in other formats such as paraffin -embedded tissue blocks. However over the past decade or so it has become clear the great majority of biospecimens stored in biorepositories may not be suited for the state-of-the art genomics, proteomics, metabolomics and other bioanalytical technologies used today to search for cancer-related biomarkers. The irreproducibility of many reported biomarkers is due at least in part to the fact that the biospecimens utilized are often procured using different techniques. Such variability can lead to significant differences in the biospecimens molecular integrity. Different methods for snap freezing surgical human tissue specimens exist, liquid nitrogen or isopentane cooled with liquid nitrogen are generally preferred for Biobanking. Freezing tissues for diagnostic and research purposes are therefore often time consuming, laborious, even hazardous, and not user friendly. In tissue banks, frozen tissue samples are stored in cryovials, capsules, cryomolds, or cryocassettes. Tissues are additionally embedded using freezing media or wrapped in plastic bags or aluminum foils to prevent desiccation. The latter method aggravates enormously further tissue handling and processing. FlashFREEZE eliminates the need for both liquid nitrogen and isopentane. Here, we describe a workflow which concurrently facilitates tissue freezing and processing for tissue banking and satisfies the qualitative demands of pathologists, cancer researchers, laboratory technicians, and tissue bankers. To overcome some of these difficulties Milestone has developed the FlashFREEZE unit to standardize the flash freezing processing step. With FlashFREEZE it is now possible to run evidence -based biospecimen protocols with full documentation needed for QA processes. FlashFREEZE combines the advantages of a state-of-the-art Stirling cooler, capable of reaching temperatures down to -80C, with the environmental and safety features of the new 3M Novec 7000 fluid. Novec is a nonflammable, nontoxic, and has low global warming potential (GWP) transfer fluid. The FlashFREEZE eliminates need for liquid nitrogen and for the toxic and flammable isopentane. Cytology touch imprints. The use of the Milestone FineFIX module in conjunction with FineFIX before staining will enhance the nuclear detail and eliminate air drying artifacts. FineFIX is an ethanol based fixation regent. FIneFIX High quality frozen sections frozen sections can be obtained by freezing the sections with PrestoCHILL and by processing the slides with FineFIX before staining. FineFIX is an Ethanol-based based patented fixation and processing reagent. Its formulation with low-toxicity additives overcomes the drawbacks commonly associate with pure Ethanol or formalin based fixatives, e.g. significant tissue shrinkage, vacuolization, pykknotic nuclei. The formalin free formulation reduces formalin exposure in the frozen section lab. Both PrestoCHILL and FlashFREEZE offer standardized temperature and time control. A USB port allows for downloading of event logs to document and download event logs. PrestoCHILL features a programmable defrost cycle to insure that the unit is frost free while maintaining optimal set temperatures during working hours. The superior quality allows the pathologist to reach an accurate diagnosis. Freezing artifacts are reduced or eliminated with PrestoCHILL. The rapid drop through the critical temperature range of 0 and -5°C reduces the size and frequency of ice crystal formation. With conventional freezing techniques, samples stay in this temperature range for a longer time, allowing the formation of ice crystals that can compress strands of fibrous tissue giving the appearance of "bubbles".