Do you have a protocol to share for re-using ultracentrifuge tubes, and have you verified that it results in contaminant-free tubes? My lab uses ultracentrifugation to isolate viruses and various extracellular vesicles, but disposing insert tubes can be rather expensive. We've considered re-use, but have some reservations: 1) resuspension of pellets is rarely 100% complete, and we can't have carryover material; 2) any substances used for cleaning would also have to be removed completely; 3) physical damage to the tubes (e.g., scratching) would make 1) and 2) more difficult.
I frequently isolate cellular suborganelles and plasma membranes employing ultracentrifuge and reuse the tubes after careful and thorough cleaning. The cleaning procedure takes time. After each use I hold each tube under running tap water for at least a minute and then dump the tube in a beaker filled with distilled water containing 10% SDS and incubate overnight at room temp. Next day I thoroughly clean inside with an applicator (long handled) fitted with cotton tips to get rid of any adhered material. Then wash again with running tap water, distilled water and finally keep the tubes submerged under double distilled water for overnight at room temp. On the following day I keep the tubes inverted in a test tube dryer rack with blotting paper at the bottom to drain the water and finally keep them straight to let them dry at room temp.
I frequently check the quality of the tubes for contaminating proteins with Na,K-ATPase and Syndecan 4 for plasma membrane, LDH for cytoplasmic and PDI for ER markers using western blots.
The ultra tubes (I use Beckman) are very expensive and our lab can't afford single use for those tubes as we use a lot almost everyday for our experimental purpose. I agree the cleaning procedure is lengthy but it is inexpensive and does not need a constant attention to prepare the tubes ready for the next use.
Reusing tubes usually leads to breakage..!! Especially if you run long hours..!! Use fresh tube which is hassle free..!!
I frequently isolate cellular suborganelles and plasma membranes employing ultracentrifuge and reuse the tubes after careful and thorough cleaning. The cleaning procedure takes time. After each use I hold each tube under running tap water for at least a minute and then dump the tube in a beaker filled with distilled water containing 10% SDS and incubate overnight at room temp. Next day I thoroughly clean inside with an applicator (long handled) fitted with cotton tips to get rid of any adhered material. Then wash again with running tap water, distilled water and finally keep the tubes submerged under double distilled water for overnight at room temp. On the following day I keep the tubes inverted in a test tube dryer rack with blotting paper at the bottom to drain the water and finally keep them straight to let them dry at room temp.
I frequently check the quality of the tubes for contaminating proteins with Na,K-ATPase and Syndecan 4 for plasma membrane, LDH for cytoplasmic and PDI for ER markers using western blots.
The ultra tubes (I use Beckman) are very expensive and our lab can't afford single use for those tubes as we use a lot almost everyday for our experimental purpose. I agree the cleaning procedure is lengthy but it is inexpensive and does not need a constant attention to prepare the tubes ready for the next use.
We re-use polyallomer tubes for CsCl density gradient ultracentrifugation of FMD virus.
It depends on the material of tubes, rotor type, and g force used.
We routinely re-use open-top, thin-walled, round-bottom polyallomer tubes running in SW28 swing-out rotors at ca. 100000 x g, 3 or 4 times without difficulty. Conical tubes of the same material (used with adapters) tend to wear out at the conical tip, but may still be re-used with care.
Sealed tubes running in vertical or near-vertical rotors at higher g forces will always undergo some deformation during the run. Do not re-use those. Also, do not re-use "ultra-clear" tubes. They are very delicate, not even resistant to high-salt conditions.
We never clean our tubes. To avoid carryover, we re-use one tube only throughout one viral preparation, for several collections from the same batch of transfected producer cells.
Depending on the type of virus you are using with the tubes, consider biosafety level issues during decontamination. Keep tubes for viral samples apart from tubes for non-viral samples. You may need official permission by your biosafety authorities, for chemical decontamination of items. In most cases this will mean, first you must prove to them that your decontamination is complete, i.e. your item is virus-free. Difficult task, when the viral vector is not able to replicate.
If you still wish decontaminate your tubes, refer to the Chemical Resistance Chart of your manufacturer. Consider only those agents to which the tube material is 100% resistant. Out of these, pick one that will best inactivate your contaminant.
Proceed with great care. Using cracked, crazed, deformed, or otherwise damaged/corroded tubes is dangerous! Gradual thinning at the bottom may not be readily visible.
Given the inevitable risk of re-using tubes after multiple rounds of chemical treatment, consider the costs of a centrifugation accident. In slightest doubt, better use new tubes.
Good luck!
We use beckman polyallomer tubes for lentiviral centrifugation and we often reuse them.
For that, after you are done with one harvest, just add 10% bleach to the tube for 2-3 min and rinse it out. Then spray with 70% ethanol a few times and rinse it. Let the tubes dry.
When you want to reuse the tubes, spray them with 70% ethanol twice. Let them dry. Then rinse them with PBS and DMEM and use them for the virus prep.
Good luck !
I use Beckman ultra tubes to purify virus. To clean the tubes first wash them with running water and then submerge the tubes into 0.1 M NaOH for two hours. Wash them again with running water and with distilled water. After that, submerge the tubes in 0.05% ortophosphoric acid for 30 minutes. Finally the tubes must be washed thoroughly with destilled water and kept inverted on a blotting paper until they be completely dry. With this procedure you do not need to use any abrassive material to clean them. Even cotton may scratch this kind of tubes after several uses.
To my experience the walls of tubes exposed to 100 000 g and more and sonicated are damaged and microscopic polycarbonate (or nitrocellulose) particles are mixed with the product of interest… especially in re-used tubes … if this trash is not a problem for further analysis it is not a big issue
We reuse Beckman ultra-centrifuge tubes to harvest lentivirus. To clean the tubes, we first submerge the tubes in 10% bleach for 30 mins, then rinse them with water completely. After that submerge the tubes into 70% ethanol for 30mins, then dry them in hood.
I agre with Alexandra, it depends on the material of tubes, rotor type, g force and matherial for ultrcentrifugation (viruses or cell particles) used.
We reuse Sorvall ultracentrifuge tubes.
If the tubes was used to harvest viruses, submerge the tubes in 10% bleach for 20-30 minits. After that, hold each tube under running tap water for 3-4 min and then fill them with destilled water and dump the tubes in a beaker with distilled water for 24 hours. Repeat this step with bidistilled water. Then spray the tubes with 70% ethanol twice. Let them dry and tubes are ready for reuse.
Use this protocol only twice.
For what is worth, some years back while doing M13 phage display we found that the slightest scratch on the inside of our polyallomer tubes resulted in phage carry-over upon reuse (even autoclaving failed to solve this particular issue). We ended up using a dedicated set of brand new tubes which were only treated by submersion into bleach, detergents, etc. and rinsed extensively (i.e. no physical scrubbing, not even with cotton tips).
They have to fit snugly into the buckets, otherwise you risk destroying the samples (and, potentially, the rotor and the centrifuge). But yes, I've done it, coincidentally it was also Beckman tubes into a Hitachi centrifuge.
I have a question for your question?
how are you supposed to use open top ultracentrifuge tubes? Are you supposed to buy the caps separately or make them yourself?
my tubes are specified for my rotor, the SW 41 ti swing bucket from beckman. However i cannot figure out weather they are supposed to be capped or not.
cheers
Luke
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