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1. Anesthetize the animal, then dissect to cannulate the exposed trachea with a catheter.
2. Cool the live lung specimen with 4°C saline.
3. (Optional) Inject a bolus of agarose gel (2%) to fill the pulmonary artery.
√ This process of filling with agarose allow for inflation of alveoli with the same
embedding material, and prevents the alveoli from collapsing during the cutting
process. Some lung specimens may be sectioned without agarose infusion, especially if
the lung specimens are from bovine or sheep animal models (because the quantity of
agarose needed for infusion exceeds several liters).
4. Using a syringe, inject a bolus of air to clear the airway.
√ Injection of air helps infused agarose reach the alveoli.
5. Select a section of the lung lobe you would like to take tissues from for sections.
6. Glue the tissue sample onto the Compresstome® specimen syringe.
7. Draw the syringe downward to bring the lung tissue core sample into the syringe.
8. Fill the syringe with 2% agarose (Sigma A-0701, low gelling point, incubated at ~37°C).
9. Cool the entire contents of the specimen syringe with the chilling block. The lung tissue sample
is now embedded in agarose. The agarose will solidify enough for stable sectioning.
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10. Load the specimen syringe onto the Compresstome® slicer.
11. The protocol is complete for preparing the lung tissue core specimen for sectioning. Proceed
from here with normal Compresstome® sectioning procedures.
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