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Homogenization of Aortic Tissue

Instrument: Bullet Blender or Bullet Blender Blue

Sample: Aortic tissue

Protocol:

  1. Cut tissue into appropriately sized pieces for analysis (10mg-300mg).  For best homogenization, cut the tissue into long, thin strips.  Pieces of tissue with a high aspect ratio will homogenize more efficiently.
  2. Strip the outer fatty later from the Aorta.  If desired, wash tissue 3x with ~1 ml PBS to remove external contaminants
  3. For samples 50 mg or greater, place the sample in Navy bead lysis kit tube.  For samples less than 50 mg, place the sample in Green bead lysis kit tube.  Alternatively, add the sample to a microcentrifuge tube then add a volume of 0.9 - 2.0 mm stainless steel bead blend equal to the mass of tissue.
  4. Add 2 volumes of buffer for every volume of sample).
  5. Close the microcentrifuge tubes securely and place them into the Bullet Blender®.
  6. Set the controls for speed 8 and run time 3 minutes. Press Start.
  7. After the run, remove tubes from the instrument.
  8. Visually inspect samples. If the homogenization is unsatisfactory, run for another two minutes at speed 10.

This protocol was adapted from: Hou CJ, Tsai CH, Su CH, Wu YJ, Chen SJ, Chiu JJ, Shiao MS, Yeh HI. Diabetes reduces aortic endothelial gap junctions in ApoE-deficient mice: simvastatin exacerbates the reduction. J Histochem Cytochem. 2008 Aug;56(8):745-52