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  • 實驗室實驗倒平板時的技巧和注意事項

    發布時間:

    2022-12-27

    作者:


    在生物實驗中相信大家都經常要倒平板,細菌培養,分子生物學實驗等等。醫藥企業中倒平板更是家常便飯,環境監測、水監測、微生物檢查等,初學者往往都倒不好,現在總結幾點小技巧供大家參考(培養基倒平板注意事項):
    一、防止倒皿時的冷凝:
    1、有經驗的都知道,手心不燙手背燙,這是一個經驗數據,此時的溫度在45度左右,最適合倒平板。 
    2、如果要連續倒多個皿,尤其是在冬天往往容易凝固,一旦凝固則需要再加熱到較高溫度才能溶化(玻璃體的特性)。所以建議此時有個50度的恒溫水浴開著,隨時把裝培養基的三角瓶放進去效果會好不少。
    二、培養皿中有冷凝水:
    一般來說,都會有幾滴,影響不大。如果過多則需要注意。大概的情況是:
    1、倒皿前將平皿最好保持在35-40度的溫度(觸手有溫熱感),
    2、保持平皿靜置環境溫度高一些。
    三、倒已冷卻的瓊脂培養基:
    對已冷卻的培養基再融化頗感頭痛心煩,因為放在電爐上烤不僅需要耐心,還得加上小心,一不留神就會沖濕瓶塞;重新放入滅菌鍋加熱吧,好是好,可又嫌費事。解決方法是用微波爐加熱,又快又好,融得徹底,用著放心。當然,裝培養基所用的三角瓶得能夠放入微波爐才行:250ml.
     
    沒有微波爐怎么辦?用電爐要加小心了,在電爐上直接加熱融化營養瓊脂時應注意在融化到上面剩最后薄薄的一層時應用小火,緩慢加熱,防止氣體沖破薄層時導致爆沸,營養瓊脂沖出三角瓶。
    四、倒平皿保持均一性:日水一次性平板圖
    傾注平皿要左三圈,右三圈的晃動,注意不要把平皿蓋沾上,并且要輕輕晃動。
    五、具體對待:
    前面幾點講了倒平皿的最適溫度保持在35-40度,但不同品牌的瓊脂質量不一樣,同樣按照國標中的配方瓊脂含量1.5%, 有些瓊脂凝固很快,有些凝固慢一些,還有環境溫度偏低,瓊脂凝固也會快一些,要根據實際情況,適當調整瓊脂的含量,在室溫較低時我們所配的培養基瓊脂 用量就偏少一些,還有在倒平板和混勻時動作要快一些,室溫實在太低,也可以把培養皿放到烘箱中適當加熱一下(40度左右)。
    六、避免氣泡:
    還有同學反映在倒平板時會產生氣泡,主要遵守以上各點,加熱不要過高,融化即可,加強操作練習,還有就是倒的時候要慢點,最好能夠貼著平皿的壁倒下去。
    來自:青島日水生物qdrishui.cn
     
    Skills and Notices of Inverting Plate in Laboratory Experiment
    In biological experiments, we believe that we often have inverted plates, bacterial culture, molecular biology experiments and so on. In medical enterprises, the inverted plate is more common, environmental monitoring, water monitoring, microbiological inspection and so on, the beginners are often not good, now summarize a few small skills for everyone to refer to:
    1. To prevent the condensation of Petri dishes:
    1, as is known to all, the palm is not hot, and it is an empirical data. At this time, the temperature is about 45 degrees.
    2. If you want to pour a lot of dishes continuously, especially in the winter, it tends to solidify easily. Once it's solidified, it needs to be reheated to a higher temperature to dissolve (the characteristics of the vitreous body). Therefore, it is suggested that a 50 degree constant temperature water bath should be opened at this time, and the effect of putting the triangle bottle into the culture medium at any time will be good.
    Two. There is condensate in the culture dish:
    Generally speaking, there will be several drops and little influence. If you have too much, you need to pay attention to it. The general situation is:
    1, the pan should be kept at a temperature of 35-40 degrees (tentacles with warm feeling) before pouring.
    2. Keep the environment at a high temperature.
    Three. The agar medium that has been cooled down:
    It is a headache for the cooled medium to melt again, because it needs not only patience on the electric stove, but also with care. It will make a wet bottle stopper without mind, and reheat it in a sterilizer. It's good, but it's hard to bother. The solution is to use microwave oven heating, fast and good, melt thoroughly, with ease. Of course, the triangular bottle used in the culture medium can be put into the microwave oven: 250ML.
    What do you do without a microwave oven? Be careful with the electric furnace. When melting the nutrient agar directly on the electric furnace, attention should be paid to the use of small fire when melting to the last thin layer above, slowly heating, preventing the gas from breaking the thin layer and causing the boiling, and nutrient agar out of the trigonometric bottle.
    Four. The Petri dish remains homogeneous.
    Pour the pan into the left three circle, shake the right three circles. Be careful not to dip the pan cover and shake gently.
    Five, specific treatment:
    The first few points said that the optimum temperature of the plate is kept at 35-40 degrees, but the quality of the different brands of agar is not the same, as well as the content of the formula agar in the national standard 1.5%, some agar solidified quickly, some solidified slowly, and the environment temperature is low, agar will solidify faster, according to the actual situation, the appropriate adjustment The agar content of the agar at room temperature is less than that of the medium agar at room temperature. It also moves faster when the plate is poured and mixed, it is too low at room temperature and can be properly heated in a oven (about 40 degrees).
    Six. Avoid bubbles:
    Students are reflected in the inverted plate will produce bubbles, mainly comply with the above points, the heating should not be too high, melting can, strengthen the operation practice, and it is the time to slow down, it is best to stick to the wall of the plate down.
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