Autoclaves, laminar flow hoods, and fridges are three of the most expensive instruments used in tissue culture labs. They all have essential roles to perform, such as an autoclave used to sterilize media, other chemicals, equipment, and tools; laminar flow hoodies used to provide a sterile environment while introducing plants in tissue culture; and a fridge is used to store all the chemicals and media at the right temperature.
Out of all these, autoclaves are a bit risky to use. And, a certain level of training is required to use the instrument. Even after receiving the training, it’s essential to take all precautions while using the instrument; and not be very casual.
This article summarizes the workings of the autoclave, how to use the equipment for your applications, and what measures you need to take while using the equipment.
How Does An Autoclave Work?
An autoclave is also known as a steam sterilizer. It uses steam pressure to kill viruses, bacteria, viruses, and spores present in equipment, tools, media, and other chemicals. The instrument has a range of applications in healthcare, biomedical, biotechnology, and other industrial operations.
The autoclave works on the principle of moist heat sterilization. High pressure inside the chamber increases the boiling point of water so that equipment can be sterilized. It also ensures that heat penetrates deeply into the machinery, resulting in irreversible loss of function and activity. When steam is moist, microbe proteins coagulate, resulting in irreversible loss of function.
Irrespective of the size and the type, the autoclave operates on the same principle. The sterilization cycle of the equipment consists of three phases, which include:
- Purge Phase: At this phase, the air is removed from the chamber. To do so, the vacuum system is given in the autoclave that replaces air with steam in the sealed chamber.
- Exposure Phase: The sterilizer drain is closed after the air is removed and the steam is continuously applied to the chamber. This results in an increase in temperature and pressure inside the chamber at the desired level. It allows the equipment to be maintained in a given physiological environment for a recommended period of time.
- Exhaust Phase: This is the last sterilization stage of the autoclave. It releases the pressure through an exhaust valve from the chamber. As a result, the ambient pressure is restored.
Why do You need To Take Precautions Using an Autoclave?
To sterilize equipment, tools, and media, it’s essential to provide high temperatures and pressure for effective sterilization. Generally, the temperature and pressure applied during the process is 121 ℃ and 15 psi for a duration of 15-20 mins.
If one doesn’t be precautious, it can cause serious harm, such as heat and steam burns, injuries to hands and arms from the door, hot fluid scalds, and bodily injury in the event of an explosion. Moreover, if biohazardous material is not packaged properly, it may cause exposure to biohazards, which may cause other issues.
Autoclave Safety Precautions
Here’s a list of some safety measures that you need to take while operating an autoclave in your lab for your safety:
- The foremost important thing is to wear Personal Protective Equipment (PPE), such as a lab coat, eye protection goggles, and heat-resistant gloves while using the instruments. It ensures your body's safety when operating the machine.
- Never seal the container tightly as it may pose an explosion risk.
- If water is flowing out of the bottom of the autoclave, do not open the door. Clogged steam lines, equipment malfunctions, or plugged drains may cause a buildup of scalding water.
- Sever superheat liquids. The superheated liquid may boil out of its containers or explode if disturbed. If workers are in a hurry to remove flasks or bottles from the autoclave, a steam explosion may occur. Therefore, don’t do any such things.
- It is not recommended to autoclave flammable, reactive, corrosive, or toxic chemicals (e.g., alcohol, chloroform, acetic acid, formalin, etc.). Lab coats contaminated with chemicals should be cleaned by an approved laundry service instead of being autoclaved.
- Inspect all glassware prior to autoclaving. Older glasswares are less stable and prone to breaking.
- Never autoclave paraffin-embedded tissue sections.
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