Culture Suspension Technique 2023

Culture Suspension Technique:-

A suspension culture, also known as a cell suspension, is a type of cell culture in which single cells or small aggregates of cells are allowed to function and multiply in an agitated growth medium.


To provide a guideline for the Preparation of culture suspension in  Microbiology Lab

 2.0 SCOPE:

2.1  The procedure is applicable to all personnel working in Microbiology Lab.


  3.1   Microbiologists/Personnel Working in Microbiology Lab

3.1.1  To follow the procedure while performing the Microbiological activities.

3.1.2   To train the new employees during on job training

  3.2     Head-Microbiology

3.2.1   To ensure compliance to the procedures

3.2.2  To ensure all the personnel are trained and adhere to the procedures


 4.1        Head Quality Assurance.

5.0 Procedure:-

 Preparation of Microbial Inoculums:

list of Micro organism used in culture suspension Activity.

Culture Suspension Technique

  1. Prepare the culture slant as per SOP for Culture maintenance.
  2. Take a loop full of culture from the working culture slant and inoculate into 10 mL Soybean Casein Digest Medium with specified organisms.

  3. Incubate for about 24 – 48 hrs. at 30 – 35 ºC for growth of organism.

  4.  After incubation period, take 1 mL from broth into 10 mL normal saline solution (0.9% w/v) and vortex it to mix uniformly. Label this solution as 10-1.

  5. Take 1 mL from 10-1 to next dilution of 9 mL of saline solution and label as 10-2 and carry out serial dilutions up to 10-9 as per below diagram.Culture Suspension Technique

  6. Pour 1 mL saline from each slant and pour with SCDA. Incubate it and observe that on which slant to obtain 100 cfu colony.

  7. Take that no. of tube for GPT test. Store that suspension at 2 – 8 ºC temp.

  8. Prepare Soybean Casein Digest Agar/Saboured Dextrose Agar as per respective SOP Media Preparation.

  9. Take 1 mL of each dilution into sterilized petri plates in duplicate and pour 15 – 20 mL of Soyabean Casein Digest Agar.

  10. Swirl the plates properly in clockwise and anticlockwise directions to mix the content properly, in such away to avoid bubble formation.

  11.  Allow the plates to solidify, incubate the plates at 30 – 35 °C for 24 – 48 hrs to Bacterial & 20 – 25 °C . for 3-5 days to fungi.

  12. Count the number of colonies in each plate and select the dilution with 10 – 100 cfu/ mL.

  13. Store the Qualified dilution of 10 – 100 cfu/ mL at 2 – 8 °C


Culture Suspension Technique

Suggested topic: Environmental Monitoring in Pharma industry.

Culture Suspension Technique

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