The BeadBeater disrupts microorganisms, single-cell preparations and pulverized plant and animal tissue with better than 95 percent efficiency. Up to 80 grams (wet weight) of cells can be processed in a typical three minute run. Relying on a unique crushing or cracking action rather than high shear forces found with a French Press or sonicator, a Teflon impeller rotating at high speed forces thousands of minute glass beads to violently collide in a special clover-leaf shaped vessel. Cells are disrupted quickly, efficiently and safely in the sealed system. The apparatus is easy to clean, has a small footprint, requires no auxiliary supplies or equipment and is essentially maintenance free. Beads settle out in seconds and are reusable. The apparatus is easy to clean, has a small footprint, requires no auxiliary supplies or equipment and is essentially maintenance free. Beads settle out in seconds and are reusable.
The BeadBeater comes complete with a 350 ml polycarbonate chamber, rotor assembly, motor base, ice-water cooling jacket and one pound of glass beads of your choice.
Like many mechanical cell disruption techniques, the Beadbeating process generates heat. In general for every minute of beadbeating there is a 10 deg rise in sample temperature. Should that be a problem, the ice-water jacket included with the BeadBeater helps keep this temperature increase under control. If you require even more temperature control, stainless steel or aluminum sample chambers are offered (see accessories).
|Homogenizer Type||Bead Mill|
|Sample Throughput||1 sample|
|Sample Volume (min)||15 ml|
|Sample Volume (max)||200 ml|
|Usage / Placement||Benchtop|
Monitoring cell lysis. The BeadBeater will disrupt over 90% of the cells in about 2-5 minutes of operation. The homogenization procedure involves cell 'cracking' action rather than high shear. After homogenization, cell membranes may still appear to be intact when viewed under a microscope. Therefore, to monitor the time course of cell breakage, rely on assay methods that measure intracellular constituents (e.g., OD260, enzyme activity, protein staining, PAGE). If the goal is to isolate intact intracelluar organelles, use the same size beads as suggested for cell disruption. Beadbeating is a very effective for this purpose. To maximize the yield of intact organelles, homogenize for a shorter period of time to get about 70% maximum cell disruption. A homogenization time study of 1, 2, 3, and 5 minutes would be instructive.
Temperature control. The homogenate will be warm after three minutes of 'BeadBeating'. When isolating proteins, membranes or organelles, cooling may be necessary. When isolating nucleic acids in aggressive extraction media containing phenol/chloroform, guanidinium salts, and/or detergents, temperature control is usually not necessary. The easiest way to minimize heating is by operating the BB, with the ice water jacket installed, for one minute and then let the homogenate sit for one minute, cycling thus until homogenization is complete. Also, consider replacing the clear polycarbonate chamber with a stainless steel closed chamber (accessory, #60801) for much better heat transfer to the ice water. For an even more stringent cooling technique see Methods in Enzymology, Vol.182, p.162-164.
Bead size selection. The correct size beads are 0.1 mm dia. for bacteria, 0.5 mm dia. for yeast, and 1.0 mm dia. or 2.5 mm dia. for chopped-up plant or animal tissue. While glass bead media is most commonly used, denser bead media is available for tough materials.
1) Fill the large, clear container or chamber 1/2 full with ice-cold beads and the rest of the volume with cells suspended in cold extraction media. Using the standard large chamber, that would be about 200 ml of beads and 200 ml of buffer containing 1-80 g wet weight of cells. Homogenizing cells at lower concentrations is okay but, in the interest of efficient down-stream purification, it may be better to keep cell concentrations high by using a Small Chamber (accessory, #110803) designed to process 15 or 50 ml of cell suspension. Place the rotor assembly on top of the container. Be sure the gray rubber gasket is in between the top lip of the container and the rotor assembly. Leave as little air as possible in the container. Holding the container and rotor assembly on one hand and the ice-water jacket (held up-side-down) in the other hand, screw them together. If temperature control is not a concern, use the Metal Ring (it looks like a thick Mason jar lid ring) to seal the filled chamber instead of the ice water jacket.
2) Invert the whole assembly, fill the ice water jacket with crushed ice and water and place the assembly on the BeadBeater motor. The bead-chain attached to the side of the motor is only used to hold down the Large Container when it is used with the Metal Ring.
3) Run the BeadBeater for about three minutes (5 min. maximum). For cooling considerations, see Temperature Control comments above.
4) The homogenate can be recovered by simply decanting. To recover the entire product, one can either pour the homogenate, beads and all, into a Buchner funnel containing filter paper, and suction the homogenate out of the beads or one can attach a glass tube with a sintered glass tip (commonly used to aerate cultures) to a side arm flask and suck out the homogenate directly from the chamber.
Wash the beads with water and then detergent. Rinse the beads repeatedly with DI or RO water and dry them overnight at 50 deg C in a shallow glass or stainless steel tray. Properly cleaned beads will be flow freely. If some of the beads cake on the sides of the drying container, repeat the cleaning process. Beads can be autoclaved or baked, if desired. If you want beads free of all nucleic acids or nucleases, soak the beads for 5 minutes in a 1:10 dilution of ordinary household bleach solution in place of the detergent wash. Beads can be reused about ten times.
Do not let a 'dirty' chamber sit around. Residual cell homogenate is corrosive and will lead to jamming and leaking of the chamber. Hand wash the plastic rotor assembly and chamber promptly.
There is a method for thorough cleaning of BeadBeater chambers. It only takes a minute more than simply washing out the intact chamber but will assure that chamber components last longer. The rotor/shaft part of the BB homogenization chamber is temporarily removed from its black plastic bushing unit. By doing so, you will remove any abrasive glass fragments that might have worked their way into the shaft and bushing area: