Let's learn the 5 I's for microbiology technique!
Inoculation
Incubation
Isolation
Inspection
Identification
Understanding these fundamental processes is essential for any researcher in the microbiology field. By using proper technique we can better understand how microbes interact with organisms and the world around us.
Inoculation
The first crucial step in the process involves taking a bacterial sample and introducing it to various sterile growth mediums. These can be solid agar media or liquid broth, both supply nutrients to microorganisms to sustain and promote growth. There is a wide range mediums that can be selective and/or differential. This means you can pick what desired organism you are hoping to grow. Differential media often uses biochemical properties of target organisms that lead to a visible change once grown. Inoculation can also be performed using different methods depending on what results you hope to see. These include streaking, spreading, or swabbing. A common technique is quadrant streaking to isolate singular species.
Incubation
After the selected microorganisms have been introduced to the media, they now must multiply. Incubating samples allows for the perfect environmental conditions for growth. Typically incubators are kept at 30-35°C with the humidity at 85-95%.
Isolation
The goal of this step is to isolate microorganisms that were previously in a mixed sample. In order to perform detailed experiments isolated colonies must be grown and separated. It's difficult to distinguish microbes since they have similar physical properties so by using techniques such as serial dilution or quadrant streaking.
Inspection
After pure and isolated cultures have been obtained, it's time to inspect and study your new findings. Observing characteristics such as size, color, and shape of the colonies is called morphology. This provides us with a clearer understanding of microbial physiology and can lead us to identify the species.
Identification
The final "i" can involve a series of tests in order to properly identify your microorganism. Through serological tests, DNA sequencing, biochemical tests, and many others your options can get narrowed down until the organisms is identified.
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