I am looking to develop an efficient high throughput tissue homogenization and extraction process to be able to accurately measure and profile androgenic steroids in small slices of mouse testes (~50mg). We have a Fisher Scientific PowerGen 125 Rotor-Stator type homogenizer that may work fine but must be thoroughly cleaned between samples to prevent cross contamination which makes throughput slow. I therefore purchased an OPS Diagnostics HTMini bead beater homogenizer with various types and sizes of grinding media in order to increase throughput and eliminate cross contamination. We also have a QSonica Q55 ultrasonic probe that reportedly works extremely well to disrupt individual cells or small groups of cells which I plan to test following one of the previous processing methods to see if it improves the extraction. I am basically starting from scratch so any suggestions would be greatly appreciated.
We've been having a lot of luck using a barocycler, see link below. We have the 2320EXT. They are not cheap, but they are great for sample limited applications. A 1 mg sample is stacks for proteomics. You can run up to 16 samples simultaneously with this model. An advantage is that each sample is in a separate tube, so there is no risk of carryover. The price of tubes and pestles are also a consideration as you'll be looking at around USD4 per sample.
The process is stupidly simple and you can start with an intact tissue and be ready for MS the same day without any issues. The equipment itself is reliable and maintenance issues are few.
Some relevant publications follow.
Hope that helps,
Peter
Shiying Shao, Tiannan Guo, Chiek Ching Koh, Silke Gillessen, Markus Joerger, Wolfram Jochum, and Ruedi Aebersold. Minimal sample requirement for highly multiplexed protein quantification in cell lines and tissues by PCT-SWATH mass spectrometry. Proteomics: 2015.
Shiying Shao, Tiannan Guo, Vera Gross, Alexander Lazarev, Ching Chiek Koh, Silke Gillessen, Markus Joerger, Wolfram Jochum, and Ruedi Aebersold. Reproducible Tissue Homogenisation and Protein Extraction for Quantitative Proteomics using MicroPestle-assisted Pressure Cycling Technology. J.Proteome Res., 2016.
http://www.pressurebiosciences.com/products/high-pressure-systems
http://pubs.acs.org/doi/abs/10.1021/acs.jproteome.5b01136
http://onlinelibrary.wiley.com/doi/10.1002/pmic.201500161/abstract
Try the FastPrep 24 from MP Bio.
It runs 24 tubes simultaneously with programmable speed/time settings. Various disposable tubes are available containing different beads for different matrices. Because the tubes are standard 2mL microtube size, they can fit in existing racks and processes
We use a tube with 6 metal beads for hair with a program lasting under 20 minutes, most of which is "resting" time, but soft tissue programs can be much shorter
We have used Barcoycler as well as Omni Bead ruptor homogenizer in our lab with good results for both the instruments. Of course the use of a tool depends on the availability (quantity and frequency) of the sample. If you have 50-100 mg sample, I would recommend use of current bead homogenizer.
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