Serum collection and sterilization
Serum collection and sterilization
Blood is collected, allowed to clot, and the serum is separated. Filter the serum through filters with progressively decreasing pore sizes. Dispense vials, package, and freeze for storage.
Operation method
Scheme 11.15 Collection and Sterilization of Serum
Principle
Blood is collected, allowed to clot, and the serum is separated. Filter the serum through filters with progressively decreasing pore sizes. Separate vials, package, and freeze for preservation. I. COLLECTION Whole blood can be collected from slaughterhouses. Blood should be collected directly from the animal, collected and placed properly before separation. Alternatively, blood should be taken directly from the live animal under the direction of a veterinarian. If the latter method is chosen, and blood is taken continuously from the same group of animals, serum can be obtained with good reproducibility, but in very small quantities and at great expense. Aseptic collection can be accomplished if done very carefully. II.Coagulation Blood placed in an airtight container at 4°C overnight forms a clot, which is called naturally coagulating serum, and which is superior to the separation of blood cells by physical methods such as centrifugation and defibrinization, since platelets release growth factors into the serum during the formation of the clot. The serum is separated from the clot and then centrifuged at 2000 g for 1 h. The precipitate is discarded. Sterilized serum is usually filtered through a sterile 0.1 μm pore size filter, but because of its high viscosity and particle content, it undergoes a series of glass fiber or other prefiltration processes before final sterile filtration. Only the final filtration requirement is sterile, with 142 to 350 mm inline disc filters or similar disposable filters (e.g., M illipak 200). The prefiltration unit can be a stainless steel filter fitted with replaceable cartridges, a disk filter or a single adhesive filter (Pall Gelman). The last one is very easy to operate but very difficult to clean and reapply.
Materials and Instruments
Sterile Filter Sterile Receiver Bottle Sterile Bottle Container Peristaltic Pump Shelf Move 1. Insert a suitable non-sterile filter membrane (if a reusable device is used) into a non-sterile filtration device (disposable glass fiber filters or 142 mm reusable filters (e.g., Pall Gelman); disposable or reusable filters with a 5 μm pore size (e.g., PallGelman Verapore); and disposable or reusable filters with a 1.2 μm pore size (e.g., Millipore Opticap); 0.45 μm pore size disposable or reusable filters (e.g., Millipore Millipak ) with a suitable non-sterile filter membrane inserted (if a reusable device is used). Caveat The above is only an example, but a range of filters and pre-filters suitable for your serum can be obtained by contacting your supplier. Disposable filters can be used if the serum is to be used only once; if the collection is to be repeated on a regular basis, it is more economical to use reusable filters for pre-filtration, and still use disposable filters for sterile filtration for added safety. Common Problems Suitable for sterilizing materials with a volume of 5 to 20 L. For more product details, please visit Aladdin Scientific website.
2. connect one or more prefilters (200 mm, 0.1 μm pore size filters (e.g., Millipak, Millipore ). 0.2 μm pore size is effective for removing bacteria and fungi, but 0.1 μm pore size can be used for removing mycoplasma), and finally a sterile disposable filtration device or a reusable filter fitted with 0.1 μm pore size membranes that are Finally, a disposable sterile filter unit or a reusable filter with a 0.1 μm pore size membrane, which is connected to a sterile receiver vessel (sterile receiver bottle with bottom outlet) by a peristaltic pump.
3. Place the pump connection into a serum container (common container used for sterility testing).
4. Turn the pump on and check the reusable filter for cracks when the pump is wet; turn the pump off and tighten the filter holder if necessary.
5. Turn the pump back on and continue filtration, checking for leaks or clogging. Increasing the flow rate will increase the speed of filtration, but may cause the filter to clog.
6. Collect equally into sterile bottles (sterile bottles covered with lids and wrapped in aluminum foil), leaving at least 20 % of the top space for thermal expansion during freezing.
7. collect samples for sterility testing at the beginning, middle and end of the process.
8. cap, mark and number the bottles.
9. re-wrap the cap of the bottle with aluminum foil up to the neck of the bottle for preservation.
10. Store at -20°C until quality control is complete.