Bullet Blender Blue from Next Advance

March 14, 2013

Overall

Performance

Ease-of-Optimization

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University of Minnesota
Dept of Medicine
Post-doctoral Fellow
Bead mill optimization using custom bead lysis kits. The bead mixtures for each kit are as follows. Maroon Lysis Kit: 100ul 0.9mm-2.0mm stainless steel beads, 5 3.2 mm stainless steel beads. Gold Lysis Kit: 50ul 0.5mm Zirconium oxide beads, 50ul 1.0mm Zirconium oxide beads. Fuschia Lysis Kit: 100ul 0.9mm-2.0mm stainless steel beads, 5 3.2mm stainless steel beads. Taupe Lysis Kit: 50ul 0.5mm Zirconium oxide beads, 50 ul 1.0mm Zirconium oxide beads, 25ul 2.0mm Zirconium oxide beads.

Company:

Next Advance

Product Name:

Bullet Blender Blue

Catalog Number:

BBX24B
Image
  • Company: Next Advance
  • Product Name: Bullet Blender Blue
  • Catalog Number: BBX24B

We are investigating differences at the proteomic level between normal and cancerous lung tissue samples. Snap frozen lung tissue must be fully homogenized and lysed in a sterile environment. Traditional mortar and pestle methods were not ideal for the high-throughput scale required for more than 200 samples. The Next Advance Bullet Blender™ Blue bead mill homogenizer offers a sterile, consistent, and efficient way to process 24 samples per run. This model comes with an air-circulator allowing the samples to remain at 4°C when the unit is kept in a cold room. Next Advance sells a variety of beads made of glass, zirconium silicate, zirconium oxide, or stainless steel. Depending on the tissue to be homogenized you can select the appropriate bead yourself or their helpful customer service reps can help you. We have processed lung tissue, mouse femurs, and fat tissue successfully with this device.

Experimental Design and Results Summary

Application

Most studies have focused on transcriptional changes when examining molecular differences between normal and cancerous lung tissue. Many of these transcriptional changes may not be reflected at the proteomic level and post-translational modifications would not be detectable at all. By examining the proteome we hope to define a series of biomarkers defining lung cancer throughout its progression providing a clearer picture of this disease process.

Starting Material

Normal and cancerous lung tissue samples

Protocol Overview

Next Advance has ready-to-use bead lysis kits available, but I chose to purchase beads in bulk and create my own lysis kits. I have optimized the bead mixture for lung tissue by adding 50 µL 0.5mm, 50 µL 1.0mm and 25 µL 2.0mm zirconium oxide beads with 100 µg of tissue and 600 µL lysis buffer. The Bullet Blender Blue™ has two prominent control knobs that modulate either the intensity or time of your homogenization run. After loading your sample, lysis buffer, and beads into a Safelock microfuge tube, you select your settings and begin homogenization. Visual inspection of the tube contents can easily verify if any intact tissue remains at which point further homogenization can easily be selected. After several trial runs, I found that two five-minute sessions at setting 12 (maximum intensity) completely homogenizes lung tissue. The tissue appears quite dissociated after the first five-minute session, but after an additional session, no identifiable structures remain.

Tips

I experimented with several mixtures of both stainless steel and zirconium oxide beads before choosing a mixture of 50 µL 0.5mm, 50 µL 1.0mm and 25 µL 2.0mm zirconium oxide beads with 100 µg of tissue and 600 µL lysis buffer. I choose this final configuration since it provided a thoroughly homogenized specimen without excessive viscosity. The stainless steel beads appear to deposit some metal dust into the mixture, which is not ideal for mass spectrometry, but may be sufficient for Western blotting or other techniques. Additionally, Trizol reagent will turn black when exposed to the stainless steel beads.

Results Summary

Lung homogenate was inspected for the presence of identifiable structures, viscosity, and total protein content. We found the ideal homogenization was achieved with 50 µL 0.5mm, 50 µL 1.0mm and 25 µL 2.0mm zirconium oxide beads with 100 µg of tissue and 600 µL lysis buffer, processed in two five-minute sessions at an intensity of 12 in a 4°C cold room.

Features Summary

The NextAdvance Bullet Blender Blue™ is a sturdy, reliable, efficient, air-cooled bead-mill homogenizer with the capacity to process 24 samples per run. The technical support department was extremely helpful and NextAdvance offers several premade bead lysis kits to facilitate efficient protocol screening. The device does generate significant noise at maximum intensity, but this is mitigated by the fact that it is housed in an insulated cold room.

Additional Notes

Next Advance sells a variety of bead sizes as well as composition materials. The Next Advance website has an extensive list of tissues and the corresponding lysis kit they recommend. The technical service department is extremely knowledgeable and can help you develop a custom protocol for your tissues as well.

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Bead mill optimization using custom bead lysis kits. The bead mixtures for each kit are as follows. Maroon Lysis Kit: 100ul 0.9mm-2.0mm stainless steel beads, 5 3.2 mm stainless steel beads. Gold Lysis Kit: 50ul 0.5mm Zirconium oxide beads, 50ul 1.0mm Zirconium oxide beads. Fuschia Lysis Kit: 100ul 0.9mm-2.0mm stainless steel beads, 5 3.2mm stainless steel beads. Taupe Lysis Kit: 50ul 0.5mm Zirconium oxide beads, 50 ul 1.0mm Zirconium oxide beads, 25ul 2.0mm Zirconium oxide beads.

Summary

The Good

Sterile, consistent, and efficient way to process 24 tissue samples per run with a high degree of customizable bead lysis kits.

The Bad

The Bullet Blender Blue is not the least expensive option unless you consider your time as having value.

The Bottom Line

The Bullet Blender Blue and Next Advance perfectly satisfy my tissue homogenization needs.

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