Sterilisation is a vital process in microbiology that ensures objects and media are free of all living microorganisms, including both vegetative forms and spores. In this article, we explore the sterilisation meaning, of sterilisation definition, and detailed sterilisation methods that are crucial in both laboratory and industrial settings. Our discussion is designed in a clear and engaging style so that even school students can grasp the concepts with ease.
Sterilisation in microbiology refers to the complete elimination of all forms of microorganisms. Unlike disinfection, which only reduces microbial numbers, sterilisation ensures that there is no viable microbe remaining on the object. This process is indispensable in creating aseptic conditions for medical instruments, culture media and many other sensitive applications.
Understanding these terms helps in recognising why various types of sterilisation are applied in different situations.
The classification of sterilisation falls into two major categories:
Physical Sterilisation Methods
Chemical Sterilisation Methods
Each method has its advantages and is chosen based on the nature of the object to be sterilised and its sensitivity to heat or chemicals.
Physical methods of sterilisation are based on utilising energy or physical forces to destroy microbial cells. Below are the key methods:
Heat is one of the most effective means to achieve sterilisation in microbiology. The sterilisation methods using heat can be divided into:
Moist Heat Sterilisation: This method uses an autoclave to produce steam under pressure at 121–134℃ with a pressure of 15 psi. The moist heat causes coagulation of proteins in the microorganisms, effectively killing them. This is the most commonly used method when a high level of sterility is required.
Dry Heat Sterilisation: When objects are sensitive to moisture, dry heat is preferred. The principle here is conduction – the heat is transferred from the surface inward, denaturing and oxidising proteins until the microbe dies. Instruments such as hot air ovens and incinerators are typically used.
Filtration is a mechanical sterilisation method that uses membranes with tiny pores to remove microorganisms from liquids and gases. The three key steps involved are:
Sieving: Trapping large particles.
Adsorption: Microbes adhere to the filter.
Trapping: Final retention of micro-organisms.
Irradiation involves exposing objects to radiation to achieve sterilisation. There are two main forms:
Non-Ionising Radiation: Ultraviolet (UV) light is used, causing the formation of pyrimidine dimers in microbial DNA, leading to replication errors and cell death.
Ionising Radiation: Gamma rays and X-rays are employed to create reactive oxygen species (such as hydrogen peroxide), which damage essential cellular components and result in microbial death.
This method employs ultrasonic waves (typically in the 20–40 kHz range) that create alternating compressive and tensile forces in a liquid. The resulting cavitation (formation and collapse of microscopic bubbles) effectively disrupts and removes microorganisms. This innovative sterilisation method is particularly useful in cleaning delicate instruments and in industrial applications.
Fractional sterilisation is used for media containing heat-sensitive components like gelatin or sugar. The process involves exposing the medium to 100℃ for 20 minutes on three successive days. The principle is that initial exposure kills vegetative cells, and any spores that germinate in the intervals are then destroyed by subsequent heating.
Chemical methods are essential when high temperatures or physical methods cannot be used. These methods are particularly important for sterilising plastic instruments and biological specimens.
In this approach, objects are placed in a closed chamber where they are exposed to gaseous chemical agents under controlled conditions. Common gases used include:
Ethylene oxide: Highly effective against all microorganisms.
Formaldehyde, nitrogen dioxide and ozone: These gases interact with microbial cell components to ensure complete sterilisation.
Liquid sterilisation involves immersing objects in a sterilising solution. Although considered less potent than gaseous methods, it is still highly useful for decontaminating surfaces and materials. Frequently used liquid agents include:
Hydrogen peroxide
Glutaraldehyde
Hypochlorite solution
In addition to the standard methods, Vedantu’s comprehensive guide on sterilisation in microbiology offers some unique insights:
Advantages and Limitations:Each sterilisation method comes with its own set of benefits and constraints. For instance, while moist heat sterilisation is highly effective, it is unsuitable for heat-sensitive items. Conversely, chemical methods can be applied to a broader range of materials but may leave chemical residues.
Emerging Technologies: Recent advances in technology have led to the development of low-temperature sterilisation systems that combine chemical and physical processes, ensuring safety for both the material and the user. Such innovative methods are being explored in fields such as tissue engineering and pharmaceuticals.
Practical Applications in Daily Life: Beyond the laboratory, sterilisation methods are used in everyday life – from ensuring the safety of packaged foods to sterilising surgical tools. Understanding these processes can help demystify everyday products and encourage informed choices about health and safety.
Environmental Considerations: With increasing focus on sustainable practices, newer sterilisation methods are being designed to minimise environmental impact. For example, research is underway to develop eco-friendly chemical sterilants and energy-efficient autoclaves.
Sterilisation is more than just a laboratory process; it is a critical component in maintaining hygiene and safety across various fields. From the sterilisation meaning and definition to the diverse methods of sterilisation—including moist heat, dry heat, filtration, irradiation, sound waves, and chemical approaches—the types of sterilisation are chosen based on the object’s sensitivity and the desired level of microbial control. Vedantu’s guide not only covers all the fundamental information but also introduces unique aspects and emerging trends, making it an essential resource for students and professionals alike.
1. What is sterilisation and how does it differ from disinfection?
Sterilisation is the process of completely eliminating or destroying all forms of microbial life, including bacteria, viruses, fungi, and their highly resistant spores, from a surface, object, or medium. It is an absolute term, meaning an item is either sterile or not. This differs from disinfection, which aims to reduce the number of harmful microorganisms to a safe level but does not typically eliminate bacterial spores.
2. What are the main methods used for sterilisation in microbiology?
Sterilisation methods are broadly classified into two main categories: Physical and Chemical methods. Each category includes several techniques chosen based on the material being sterilised.
3. How do moist heat and dry heat sterilisation work, and when is each type used?
Both are heat-based methods but work on different principles. Moist heat sterilisation, typically done in an autoclave, uses steam under pressure (e.g., 121°C at 15 psi) to kill microbes by coagulating their essential proteins. It is fast and effective for thermostable items like surgical instruments and culture media. Dry heat sterilisation, using a hot air oven, employs higher temperatures (e.g., 160-180°C) for longer durations to kill microbes through oxidation. It is used for materials that are sensitive to moisture, such as powders, oils, and certain glassware.
4. Why can't heat-based sterilisation be used for all materials, and what are the alternatives?
Heat-based methods cannot be used for all materials because high temperatures can damage or destroy heat-sensitive (thermolabile) items. For instance, many plastics, electronic components, and biological solutions like vaccines or certain antibiotics would be ruined. In such cases, low-temperature sterilisation methods are used as alternatives. These include:
5. What are some real-world examples of sterilisation in healthcare and daily life?
Sterilisation is a critical process with many real-world applications. In healthcare, it is essential for preparing surgical instruments, needles, and implantable medical devices to prevent infections. In the food industry, it is used to ensure the safety and shelf-life of canned foods. In a laboratory setting, scientists sterilise culture media, glassware, and equipment to prevent contamination of experiments. Even the needles used for tattoos and piercings must be properly sterilised.
6. How does irradiation achieve sterilisation without using heat?
Irradiation achieves sterilisation by using energy to damage the fundamental components of microorganisms, primarily their DNA. Ionising radiation (like gamma rays) creates highly reactive free radicals within the microbial cell, which damage DNA and proteins, leading to cell death. Non-ionising radiation (like UV light) is less penetrating but causes adjacent pyrimidine bases in the DNA to bond together, forming dimers. These dimers disrupt DNA replication and lead to the microbe's death. This entire process occurs without significantly raising the temperature of the item.
7. What is fractional sterilisation and why is it used?
Fractional sterilisation, also known as Tyndallisation, is a method used for sterilising media that contain heat-sensitive components like sugars or gelatin. The process involves heating the medium to 100°C for about 20 minutes on three consecutive days. The first heating kills vegetative bacterial cells. During the incubation period between heatings, any resistant spores germinate into new vegetative cells, which are then killed during the subsequent heating cycles. This ensures sterility without damaging the heat-sensitive ingredients.