Скачать с ютуб Sample Preparation Tutorial Part I: How to Freeze Your Tissue Sample To Ensure High-Quality Results? в хорошем качестве

Sample Preparation Tutorial Part I: How to Freeze Your Tissue Sample To Ensure High-Quality Results?

Are you having trouble freezing your tissue sample for the Molecular Cartography workflow? Here are the tips and tricks to help you properly freeze the tissue before sectioning. For more technical questions, please get in touch with our regional Customer Technology Advisor or send us an email at [email protected]. We are always happy to help if you have any issues during tissue preparation. Full transcript: Hello, my name is Jeroen, and I'm a Customer Technology Advisor with Resolve Biosciences. Today, I will be showing you how to freeze your tissue sample to ensure the highest quality for your Molecular Cartography experiments. Let's get started. You will need a box with dry ice, a beaker, and a bottle of isopentane. Isopentane is much more suitable than liquid nitrogen to preserve tissue morphology in larger samples. Keep in mind to safety measures when working with isopentane and proceed with the protocol under a fume hood. For more information, please consult the safety sheet supplied by the isopentane manufacturer. Fill the bottom of the beaker with isopentane to around three or four centimeters high. Meanwhile, prepare pieces of aluminum foil and label them with sample information. Keep them cold on dry ice. These will be needed to store the samples after freezing. Now back to our isopentane setup. Make sure to stir the liquid regularly to ensure a homogeneous temperature. For convenience, we use a thermometer for this to simultaneously measure the cooling progress. Ensure that the isopentane is evenly cooled to approximately -55 degrees Celsius. To freeze the sample, you have two options depending on the sample size. For a large sample, such as a mouse brain, for example, you can place the freshly dissected or fixed tissue directly in isopentane. A great trick is to use a weighing tray to make it easy to retrieve the tissue. First, make a small snippet in the tray using scissors, which will help to decant the isopentane. Place a tissue on the tray and submerge it in the isopentane for about one minute. Retrieve the tissue and decant the isopentane from the tray. Wrap the tissue in a pre-cooled and labeled piece of aluminum foil and store the wrapped tissue at -80 degrees Celsius until sectioning. For small samples, we advise to use cryomold and OCT to freeze the sample. Place a drop of OCT at the bottom of the cryomold. Place the tissue with your region of interest to section at the bottom and fill the rest of the cryomold with OCT. Submerge the mold in isopentane for about two minutes until the OCT is completely white. Instead of using a beaker, it can be easier to submerge the sample in a reservoir to keep the OCT in place. Retrieve the cryomold and completely wrap it in a pre-cooled and labeled piece of aluminum foil. Store at -80 degrees Celsius until section. #spatialtranscriptomics #spatialbiology #spatialgenomics #tissuefreezing