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SOP

Colony Counting

Procedure for counting microbial colonies on suitable culture media and calculating quantitative results using validated criteria.

Last verified 1 month ago 2 min read Microbiology Quantitative Microbiology #cfu #colony-count #Enumeration
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Document Control
Version
1.0
Effective
Aug 31, 2026
Reviewed
Aug 28, 2026
Department
Microbiology
Principle

Under appropriate culture conditions, viable organisms form visible colonies that can be counted within the validated quantitative range of the method.

Purpose

To standardize counting and calculation of colonies for quantitative microbiological examinations.

Scope

This procedure applies to trained microbiology laboratory personnel working within an approved quality-management and biosafety system.

Principle

Under appropriate culture conditions, viable organisms form visible colonies that can be counted within the validated quantitative range of the method.

Specimen / Test Material

Appropriately inoculated culture plates from validated quantitative procedures.

Equipment and Materials

  • Suitable culture plates
  • Colony counter or manual counting aid
  • Marker or approved recording system
  • Calculator or validated laboratory information system

Procedure

  1. Confirm plate identification and culture integrity.
  2. Select plates meeting the validated counting criteria.
  3. Count colonies using the approved method.
  4. Record counts and dilution factors.
  5. Calculate the final result using the validated formula.
  6. Apply laboratory rules for plates outside the validated counting range.

Quality Control and Quality Assurance

  • Verify counting criteria.
  • Check calculation accuracy.
  • Review unusual counts or contamination patterns.

Reporting

Report quantitative results in the approved units and include relevant limitations.

Safety

Use risk-based biosafety practices, appropriate personal protective equipment and approved decontamination and waste-disposal procedures. Treat clinical specimens and cultures according to the laboratory's biological risk assessment.

References

  • World Health Organization (WHO): Toolkit for Standard Operating Procedures for Bacteriology, Mycology and Antimicrobial Susceptibility Testing.
  • WHO: Laboratory Biosafety Manual, 4th edition.
  • WHO guidance on collection, packaging and transport of infectious substances.
  • EUCAST methodology and quality-control guidance for antimicrobial susceptibility testing, where applicable.
  • Current locally approved laboratory procedures and validated manufacturer instructions.

Frequently Asked Questions

No. Use plates that meet the validated method's acceptance and counting criteria.
They are required for correct calculation of the original concentration.