Cryosectioning Recommendations

How to cut Tissue Samples

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Sample Preparation Recommendations

CARTANA recommends tissue thickness of 10 +/- 5 µm, collected on SuperFrost® microscope slides.

1. Mount the Optimal Cutting Temperature (OCT) compound embedded sample on the specimen chuck.

  • Place the sample in the cryostat and leave it for 15 minutes to equilibrate.
  • Adjust sample orientation.

2. Cut and discard few sections to have a clean and straight surface on the tissue.

  • Cut a section and flip the tissue upside down to avoid curling.
  • Remove the folds and wrinkles present on the cut sections with a clean brush.
  • Make sure you do that by touching only the OCT around the section and not the tissue.

3. After you collect the section, leave the slide at room temperature until the section is dry (~1 minute).

  • Quickly transfer the slide in the cryostat or store at -80°C.
  • Repeat the same procedure with the following slide.

You can place several sections (corresponding to the approximate size of the SecureSeal chamber format you will use) on one slide with few mm distance between them. You can freeze your glass slide at -80°C for later processing or continue directly with the CARTANA HS Library Preparation Kit protocol to prepare the library for In Situ Sequencing.

It is recommended to work in an RNase free environment to avoid degradation of RNA molecules. Always use fresh RNAse free reagents for sample preparation.

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