常見培養(yǎng)基質(zhì)量控制問題匯總
發(fā)布時間：
2022-12-24
作者：
上海微生物培養(yǎng)基
常見培養(yǎng)基質(zhì)量控制問題匯總
1、平板涂布和平板劃線培養(yǎng)基表面水分過多，菌落蔓延如何解決？
對于采用表面接種形式培養(yǎng)的固體培養(yǎng)基，應(yīng)先對瓊脂表面進(jìn)行干燥：揭開平皿蓋，將平板倒扣于烘箱或培養(yǎng)箱中（溫度設(shè)為25℃～50℃）；或放在有對流的無菌凈化臺中，直到培養(yǎng)基表面的水滴消失為止。注意不要過度干燥。商品化的平板瓊脂培養(yǎng)基應(yīng)按照廠商提供的說明使用。
2、滅菌后的培養(yǎng)基的存放時間，保存方式？
瓊脂培養(yǎng)基的融化
將培養(yǎng)基放到沸水浴中或采用有相同效果的方法（如高壓鍋中的層流蒸汽）使之融化。經(jīng)過高壓的培養(yǎng)基應(yīng)盡量減少重新加熱時間，融化后避免過度加熱。融化后應(yīng)短暫置于室溫中（如2 min）以避免玻璃瓶破碎。
融化后的培養(yǎng)基放入47℃～50℃的恒溫水浴鍋中冷卻保溫(可根據(jù)實際培養(yǎng)基凝固溫度適當(dāng)提高水浴鍋溫度)，直至使用，培養(yǎng)基達(dá)到47℃～50℃的時間與培養(yǎng)基的品種、體積、數(shù)量有關(guān)。融化后的培養(yǎng)基應(yīng)盡快使用，放置時間一般不應(yīng)超過4 h。未用完的培養(yǎng)基不能重新凝固留待下次使用。敏感的培養(yǎng)基尤應(yīng)注意，融化后保溫時間應(yīng)盡量縮短，如有特定要求可參考指定的標(biāo)準(zhǔn)。
傾注到樣品中的培養(yǎng)基溫度應(yīng)控制在約45℃左右。
平板的制備和儲存
傾注融化的培養(yǎng)基到平皿中，使之在平皿中形成厚度至少為3 mm（直徑90 mm的平皿，通常要加入18 mL～20 mL瓊脂培養(yǎng)基）。將平皿蓋好皿蓋后放到水平平面使瓊脂冷卻凝固。如果平板需儲存，或者培養(yǎng)時間超過48 h或培養(yǎng)溫度高于40 ℃，則需要傾注更多的培養(yǎng)基。凝固后的培養(yǎng)基應(yīng)立即使用或存放于暗處和（或）5℃±3℃冰箱的密封袋中，以防止培養(yǎng)基成分的改變。在平板底部或側(cè)邊做好標(biāo)記，標(biāo)記的內(nèi)容包括名稱、制備日期和（或）有效期。也可使用適宜的培養(yǎng)基編碼系統(tǒng)進(jìn)行標(biāo)記。
將倒好的平板放在密封的袋子中冷藏保存可延長儲存期限。為了避免冷凝水的產(chǎn)生，平板應(yīng)冷卻后再裝入袋中。儲存前不要對培養(yǎng)基表面進(jìn)行干燥處理。
對于采用表面接種形式培養(yǎng)的固體培養(yǎng)基，應(yīng)先對瓊脂表面進(jìn)行干燥：揭開平皿蓋，將平板倒扣于烘箱或培養(yǎng)箱中（溫度設(shè)為25℃～50℃）；或放在有對流的無菌凈化臺中，直到培養(yǎng)基表面的水滴消失為止。注意不要過度干燥。商品化的平板瓊脂培養(yǎng)基應(yīng)按照廠商提供的說明使用。
Summary of common media quality control problems
1. How to solve the problem of colony spread when there is too much water on the surface of plate coating and plate marking media?
For solid medium using surface vaccination form, should be carried out on the AGAR surface drying: uncover plate cover, will buckle the tablet in the oven or in the incubator (temperature is set to 25 ℃ to 50 ℃); Or put it in a sterile purification stand with convection until the droplets on the surface of the culture medium disappear. Be careful not to overdry. The commercialized plate AGAR media shall be used in accordance with the instructions provided by the manufacturer.
2. Storage time and preservation method of sterilized media?
Melting of AGAR medium
Melt the medium in a boiling water bath or by using the same method as laminar steam in a pressure cooker. The high pressure medium should minimize the reheating time and avoid excessive heating after melting. After melting, the glass bottle should be placed at room temperature (eg 2 min) for a short time to avoid breakage.
Melted medium in 47 ℃ to 50 ℃ constant temperature water-bath water in cooling heat preservation (according to actual medium solidification temperature raise the temperature of water bath pot), until the use, medium to 47 ℃ ~ 50 ℃ time associated with the variety, size, quantity of medium. The melted medium should be used as soon as possible and generally should not be placed for more than 4 hours. The unused medium cannot be re-solidified for another time. Especially for sensitive media, it should be noted that the time of heat preservation after melting should be shortened as far as possible. If there are specific requirements, the specified standards can be referred to.
Poured into the sample medium temperature should be controlled at about 45 ℃ or so.
Plate preparation and storage
Pour the melted medium into the plate to form a plate at least 3 mm thick (90 mm in diameter, usually 18 mL ~ 20 mL AGAR medium). Cover the plate and put it on the horizontal plane to cool the AGAR. If tablet should be stored, or more than 48 h of incubation time or the temperature higher than 40 ℃, then you need to devote more of the culture medium. After solidification of media should be used immediately or stored in the dark and (or) 5 ℃ + 3 ℃ refrigerator sealing bag, in order to prevent the medium components change. Mark the bottom or side of the plate with a name, preparation date, and/or expiration date. The appropriate medium coding system can also be used for labeling.
Keep the inverted plate in a sealed bag and refrigerate for longer storage. In order to avoid condensation, the plate should be cooled and then put into a bag. Do not dry the surface of the medium before storage.
For solid medium using surface vaccination form, should be carried out on the AGAR surface drying: uncover plate cover, will buckle the tablet in the oven or in the incubator (temperature is set to 25 ℃ to 50 ℃); Or put it in a sterile purification stand with convection until the droplets on the surface of the culture medium disappear. Be careful not to overdry. The commercialized plate AGAR media shall be used in accordance with the instructions provided by the manufacturer.