Culturing with a Bunsen burner

A sterile work area for microbiological culturing can be created around a Bunsen burner or in a biosafety cabinet. The techniques used to work aseptically near a Bunsen burner and in a biosafety cabinet are similar.

The Bunsen burner creates a radial sterile field around it. Items can also be sterilized by passing them through the flame. This allows for the use of reusable equipment such as wire loops and glass spreaders. The key steps to correct aseptic technique using a Bunsen burner are listed below.

Preparing the sterile work area

• Decontaminating the work surface with ethanol
• Light the Bunsen burner and adjust to produce a roaring blue flame
• Place only necessary materials in the sterile work area and arrange them so the updraft that is creating the sterile field is not disturbed

Culturing technique

• Minimize movements that can disturb the sterile field around the Bunsen burner
• Hold any lids and closures in the hand in the sterile field rather than placing them on the work surface
• Sterilize any equipment, including the necks of bottles, by holding it in the flame
• Use either a new disposable item (such as pipette tips) for each action or sterilize reusable equipment (such as glass spreaders) before and after each action

Decontamination and waste disposal

• Sterilize all reusable equipment, close all containers and dispose of all disposable items in the biological waste bin
• Turn off the Bunsen burner and decontaminate the work surface with ethanol
• Disinfect your hands

Bunsen Burner safety measures

• Tie back long hair and avoid loose items of clothing near the flame
• Gloves should never be worn, instead, wash hands thoroughly before and after working with microbiological samples or disinfect hands with hand sanitizer
• Ethanol should only be used while the Bunsen burner is off