Can feces be stored at -80°C for in vitro fermentation?
Feces can be stored at –80°C for in vitro fermentation, but it is recommended to add a cryoprotectant and use it as soon as possible. If high microbial viability is required, fresh samples should be prioritized. During storage and thawing, anaerobic conditions should be maintained as much as possible to reduce microbial loss.
1. Pay attention to storage and resuscitation methods
1. Feasibility of storage
Fecal samples can be frozen at –80°C for short-term use (1-3 months) in in vitro fermentation, but freezing can affect the viability of some live bacteria and the structure of microbial communities, particularly those sensitive to anaerobic conditions.
2. Add cryoprotectant
To preserve microbial viability, it is recommended to add a cryoprotectant (such as 10% glycerol) and mix well before freezing, which is more conducive to subsequent resuscitation.
3. Operation during use
Thawing should be rapid and preferably conducted under anaerobic conditions, such as in an anaerobic chamber or pouch, to minimize the damage of oxygen exposure to the microbiota.
2. Attention
1. Freezing feces may cause slight changes in community composition and activity, and is less stable and reliable than fresh samples.
2. If your experiment is highly sensitive to microbial metabolic activity and fermentation products (such as SCFAs, gases), fresh samples should be prioritized.
3. If it is only used for community composition analysis (such as 16S sequencing), storage at –80°C is the standard method, with minimal impact.
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