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>
No Cross Contamination
The samples are homogenized (or mixed)
non-invasively. The sample tubes are kept closed during
agitation, as the samples are processed. There are no probes to
clean between samples.
> Samples Stay Cool
The instrument uses very little power due to the
unique, patented design. It uses a small DC motor to agitate the
individual tubes, not ultrasonics. Also, because the motor
does not need to agitate any heavy platforms or plates, it is small and
will last for years.
>
Convenient
to Use
Simply place your cartilage and other tissue samples or
cell cultures and some beads in standard high quality polypropylene
tubes, and load the tubes into the Bullet Blender™.
Set the duration (typically a few minutes) and speed
(vigorousness). There are no probes to clean. And it is
quieter than a sonicator and does not heat up your samples more than a
few degrees!
>
Risk Free
Purchase
The Bullet
Blender™ comes with a 30-day
money back guarantee and a two year warranty, with a three year
warranty on the motor. The simple, reliable design enables the Bullet Blenders™ to sell for a fraction
of the price of ultrasonic or other agitation based instruments, yet
provides an easier, quicker technique.
> Established Protocols
Protocol for Cartilaginous Tissue Homogenization in the Bullet Blender™
The protocol described in this document is for the use of the Bullet Blender™ for the
homogenization of cartilage (from a variety of animals). Note that the time and speed
settings, and digestion parameters may differ due to the variation in consistency/texture
of tissue from species to species. This protocol does not specify a particular
homogenization buffer - you may choose which is most appropriate for your downstream
application (nucleic acid isolation, protein extraction, etc.).
Materials Required:
cartilage, aspirator, Bullet Blender™, homogenization buffer, pipettor,
testicular hyaluronidase (H-3506, Sigma Chemical, St. Louis, MO),
trypsin-EDTA (0.25%, Invitrogen, Carlsbad, CA), collagenase type II
(CLS2, Worthington, Lakewood, NJ), Next Rocker platform rocke r,
microcentrifuge tubes, and 0.9-2.0mm stainless steel bead blend (part number SSB14B)
Instructions:
- Dice cartilage tissue (20-100mg) into small pieces (~2mm squares) and place into
a microcentrifuge tube.
- OPTIONAL: Wash tissue 3x with ~1mL PBS. Aspirate. NOTE: This step removes
external contaminants (blood, connective tissue, etc.).
- Add 1mL hyaluronidase to sample and incubate (15 minutes at 37°C, on Next
Rocker). Centrifuge at 1000g for 5 minutes. Aspirate supernatant.1
- Add 1mL trypsin-EDTA to sample and incubate (30 minutes at 37°C, on Next
Rocker). Centrifuge at 1000g for 5 minutes. Aspirate supernatant.1
- Add 1mL collagenase, type II (2 to 4 hours at 37°C, on Next Rocker). Centrifuge at
1000g for 5 minutes. Aspirate supernatant.1
- Add a mass of the stainless steel bead blend equal to 3x the mass of tissue. One
scoop of beads ≈
220mg.
- Add 0.1mL to 0.6mL homogenization buffer (2 volumes of buffer for every mass of
tissue).
- Close the microcentrifuge tubes.
- Place tubes into the Bullet Blender™.
- Set controls for SPEED 10 and TIME 5 minutes. Press Start.
- After the run, remove tubes from the instrument.
- Visually inspect samples. If homogenization is unsatisfactory, run for another five
minutes at the SPEED 10.
- Proceed with your downstream application.
SAFETY NOTE!!!
When using a centrifuge to separate your homogenate from
the debris and beads, make sure your tubes are balanced.
Notes
This protocol is a modified version from the publication "Cartilage Tissue Engineering for
Laryngotracheal Reconstruction: Comparison of Chondrocytes from Three
Anatomic Locations in the Rabbit" Tissue Eng. 2007 April ; 13(4): 843–853.
1 OPTIONAL: You may wash the sample with 1mL PBS after the digestion.
Centrifuge at 1000g for 5 minutes. Aspirate supernate. Continue with
protocol.
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