Name: autoclaving lab materials safely
Uploaded: 2023-04-30T06:57:36.000Z
Duration: 6 min 34 s
Channel: JoVE

K12 Standards

Texas Essential Knowledge and Skills

High School

Specialised Topic in Science

112.51.c.1.

112.51.c.1.C.

Proper Use of Autoclaves

1. Wear Proper Personal Protective Equipment (PPE)
The high pressure and temperature operating conditions encountered during autoclaving pose potential heat burn or explosion risks. Appropriate PPE includes a laboratory coat, safety glasses, heat resistant gloves, and closed-toe shoes; these must be worn when loading and unloading materials from the autoclave. When dealing with liquid samples, wear an apron and face shield as additional PPE.
2. Preparation of Materials to be Loaded
<ol>
	<li>Identify Autoclave Compatible Material and Autoclave Incompatible Material 
	Although autoclaving represents a commonly used and economic method for decontamination, not all materials are suitable for autoclaving due to the high pressure and temperature working conditions. In general, all materials that will evaporate or melt under the high-pressurized steam are not compatible with autoclaving. This includes materials like organic solvents, poorly heat resistant plastic such as polystyrene, polyethylene, and metals excluding stainless-steel. Never autoclave flammable, corrosive, toxic, reactive, or radioactive chemicals. Materials compatible for autoclaving include Pyrex or Type I borosilicate glass, polypropylene, polycarbonate, gloves, stainless steel, pipette tips, paper (put inside autoclave bags), and media solutions.</li>
	<li>Packaging Material 
	After identifying autoclave-compatible materials, suitable packaging is also important since it ensures safety and efficient autoclaving. Here are the basic principles when packaging materials:
	<ol>
		<li>Always use a secondary container made of polypropylene or stainless steel.</li>
		<li>For liquid samples, fill the vial around &#189; full and loosen the caps or use vented closures. Never fill the containers over 2/3 full with the cap fully tightened.</li>
		<li>Inspect the glass vessel to make sure there are no cracks. If cracks are identified, dispose of glassware in a proper receptacle. Never autoclave a known broken glass vessel.</li>
		<li>Always use autoclave-compatible bags to package waste. The bags should have indication tapes on it and should be opened at the ends before loading to allow steam inside the bag.</li>
		<li>Never overload the bags or container to allow sufficient steam flow for complete sterilization. Leave space between items.</li>
		<li>Don&#39;t mix incompatible materials.</li>
		<li>Never place sharp items in the waste bag.</li>
	</ol>
	</li>
</ol>
3. Load Material
<ol>
	<li>Check the autoclave&#39;s interior to make sure there is no potential hazard left from the previous user.</li>
	<li>Clean the drain before loading materials to allow good circulation.</li>
	<li>Make sure the bags do not touch the interior walls of the autoclave to avoid melting.</li>
	<li>Liquids and dry materials require different cycles and need to be autoclaved separately.</li>
	<li>Close the door firmly.</li>
</ol>
4. Operation of Autoclave
<ol>
	<li>Only trained personnel should be allowed to operate autoclaves. Refer to the equipment manual if needed, since operation protocols vary among different autoclaves.</li>
	<li>After making sure the autoclave door is firmly closed, set the autoclaving temperature to 121 &#176;C and maintain a pressure of at least 15 psi.</li>
	<li>To set the autoclave time and cycle, the following factors need to be considered:
	<ol>
		<li>Equipment recommendations from the manufacturer&#39;s manual.</li>
		<li>The type of loaded materials inside: dry or liquid.</li>
		<li>The amounts of material(s) loaded.</li>
		<li>Shape and size of container used.</li>
		<li>Heat conductivity of container and materials.</li>
		<li>Purpose of decontamination.</li>
	</ol>
	</li>
</ol>
5. Unloading of Autoclaved Material(s)
<ol>
	<li>Wear proper PPE as described above.</li>
	<li>Make sure the temperature and pressure of the autoclave have returned to a safe range.</li>
	<li>Stand behind the door and carefully open the door to release any leftover steam inside the autoclave into the room.</li>
	<li>Allow the materials inside the autoclave to stand for 10 min to release any steam or hot air trapped inside the material.</li>
	<li>Do not shake any liquid material while removing it from the autoclave. If necessary, label the material as hot to avoid heat burns to an individual who may be potentially unaware of the fact that the material has recently been autoclaved.</li>
	<li>Wait for the material to cool down to room temperature before transporting within the laboratory or outside the laboratory. In both instances, be sure to utilize appropriate containment for transporting.</li>
	<li>Record the details of the materials and autoclaving conditions in a user&#39;s log.</li>
</ol>

Compared to chemical or radiation decontamination methods, autoclaving represents the most economic and practical method in the laboratory. However, to ensure safety (avoiding melting, heat burns, or explosion), identify autoclave-compatible materials and practice proper packaging, loading, and unloading. Ensuring sufficient decontamination efficiency requires container or bag materials and enough space between materials within the autoclave. Maintenance and inspection of the autoclave on a regular schedule are required ...

proper use of autoclaves

Robert M. Rioux &amp; Zhifeng Chen, Pennsylvania State University, University Park, PA
Autoclaving is one of the most commonly used methods in the laboratory for the purpose of decontamination. The most common items decontaminated with an autoclave are those that contact biological samples (typically those containing microorganisms). An autoclave works by utilizing pressurized high temperature steam to kill microorganisms present in the loaded materials. Sufficient steam flow and heat transfer are essential for highly efficient autoclaving, which are the key principles to consider when packaging materials. Safety needs to be considered when working with an autoclave due to the high pressure and temperature employed therein, which also sets limits on which materials are compatible and may necessitate special attention when packaging, loading, and unloading materials.

Robert M. Rioux &amp; Zhifeng Chen, Pennsylvania State University, University Park, PA
Autoclaving is one of the most commonly used methods in the ...

高压灭菌是实验室中最常用的去污方法之一。许多科学家，尤其是那些处理生物样品的科学家，需要消毒的设备和介质，而高压灭菌器提供了一种快速且经济的材料净化方法。它们使用高温高压下的蒸汽来杀死微生物。但是，由于这些极端条件，必须谨慎使用它们。该视频将说明高压灭菌器的工作原理以及如何正确使用它们。在高压釜中，材料在至少 15 p.s.i 的压力下用蒸汽加热到 121 摄氏度的温度。至少 30 分钟后，这些情况会使细胞脱水并杀死细胞。高压灭菌器可以对液体和固体材料进行灭菌，但必须考虑不同的作设置。液体需要比固体更长的时间来排出蒸汽，以防止闪蒸。此外，大量液体材料需要更长的时间才能加热到正确的温度。并非所有材料都能承受高压釜内的温度。因此，切勿对任何会熔化、燃烧、易爆或会蒸发的液体进行高压灭菌。此外，不要高压灭菌有毒或放射性化学品。现在，我们将向您展示如何为高压蒸汽灭菌器制备样品以及如何作高压蒸汽灭菌器。首先，选择您将进行高压灭菌的材料，并且仅使用兼容的容器和仪器。耐热玻璃、硼硅酸盐玻璃、不锈钢、聚丙烯和聚碳酸酯可用于高压灭菌。其他常见的实验室材料，如移液器吸头和盒子，也兼容。纸张可以在兼容高压灭菌器的袋子中高压灭菌。固体废物也可以用合适的袋子进行高压灭菌。在这个袋子里留出空间，让蒸汽循环并系住两端。最后，在高压灭菌之前，确保在袋子上开一个小口，让蒸汽进入。对于水性介质和其他兼容的液体样品，请使用没有任何裂纹的完整容器。将容器装满大约一半，不要超过三分之二。要从液体容器中排出气体，请松开盖子。如果没有盖子，请用铝箔松散地盖住，以防止高压灭菌材料受到污染。将所有材料放入聚丙烯或不锈钢制成的辅助容器中。将要高压灭菌的固体和液体材料分开。在材料之间留出空间，不要将容器装得太满。最后，在所有材料上贴上一条高压灭菌胶带。该胶带具有温度敏感指示器，如果达到足够的温度，该指示器将变为黑色。使用高压灭菌器时，请佩戴适当的个人防护设备，以防止高温和高压风险。这包括标准的 PPE 安全眼镜、实验室外套、露趾鞋和耐热手套。处理液体样品时，请使用围裙和面罩。在高压灭菌器上，首先打开门并检查内部是否有任何危险。分别加载液体和干燥材料，因为它们需要不同的高压灭菌循环。为防止熔化，请确保材料不要接触墙壁。通过在材料之间保持足够的间距来确保良好的热量流动。最后，关上并密封门。不同的高压蒸汽灭菌器有不同的作说明。可以通过手动设置温度和时间或从预设列表中进行选择来设置循环。通常，请使用针对您要净化的材料类型的推荐循环。固体和液体以及不同数量的材料和废液存在多种循环。当您运行循环时，请确保温度至少达到 121 摄氏度，压力至少为 15 p.s.i.循环结束后，确保压力已恢复为零，并且高压灭菌器已冷却。小心地打开门，让剩余的蒸汽排出。然后，让材料静置 10 分钟以释放任何热气体。高压灭菌胶带现在应该有黑色条纹或上面写着高压灭菌，尽管这并不能保证已经进行了灭菌。穿戴适当的个人防护装备，去除材料，确保不要摇晃任何液体样品。在日志中记下您的使用情况，包括物质类型和遇到的任何特殊情况。最后，将材料运输到安全区域或层流罩中冷却。您刚刚观看了 JoVE 对高压灭菌器的介绍。您现在应该了解哪些材料可以高压灭菌、如何制备它们以及如何安全地使用仪器。感谢观看！

Education

教育

JoVE Science Education

Basic Biology

基础生物学

Lab Safety

实验室安全

正确使用高压釜

autoclaving lab materials safely

Autoclaving lab materials safely is an important part of decontamination in the laboratory. This method is commonly used for items that have contacted biological samples, especially those that may contain microorganisms.

An autoclave cleans materials with pressurized steam at a high temperature. The steam and heat work together to kill microorganisms on the loaded items. For this reason, effective autoclaving depends on good steam flow and strong heat transfer.

Packaging matters because it affects how well steam reaches the materials. Safe use also matters because an autoclave uses high pressure and high temperature. These conditions limit which materials can go inside and require care when packaging, loading, and unloading items.

Autoclaving Lab Materials Safely

proper personal protective equipment

Choosing the Right Lab PPE

Choosing the right lab PPE is a key part of working safely in a laboratory. PPE, or personal protective equipment, helps reduce risk when hazards are present. Laboratory hazards can be many and varied. The right choice of PPE helps make the lab a safe place to work. Students should match their protective gear to the tasks and conditions in the lab. This topic focuses on selecting proper protection for the situation at hand. Wearing the correct PPE supports safer lab work and helps limit...

emergency eyewash and shower stations

Laboratory Chemical Spill Response Stations

Laboratory chemical spill response stations help protect people who are exposed to injurious or corrosive substances. In workplaces and school labs, OSHA requires emergency eyewash and shower stations to be easy to reach when these hazards are present. These stations are used after an accident that spills a harmful chemical on the body or splashes it into the eyes. The eyewash station helps rinse the eyes, while the shower station is used for chemicals on the skin or clothing. They are meant...

electrical safety

Preventing Lab Electric Shock and Fires

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working with centrifuges

Centrifuge Safety and Sample Separation

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working with hot and cold sources

Safe Use of Lab Heat and Cold Sources

Safe use of lab heat and cold sources is an important part of laboratory work. Extreme temperatures are common in many lab tasks. They can involve both high heat and very low temperatures. Bunsen burners and hot plates are widely used in research labs and industry. A Bunsen burner is a gas burner that gives a direct flame. A hot plate is an electric device that warms containers without an open flame. Because these tools reach high temperatures, users need to follow safe handling procedures. <br...

guidelines in case of a laboratory emergency

guidelines-in-case-of-a-laboratory-emergency

Lab Spill and Accident Response Basics

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chemical storage: categories, hazards and compatibilities

chemical-storage-categories-hazards-and-compatibilities

Safe Chemical Labels and Storage Basics

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safe handling of mineral acids

Mineral Acids in the Lab

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handling chemical spills

Chemical Spill Safety in the Lab

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Autoclaving lab materials safely is an important part of decontamination in the laboratory. This method is commonly used for items that have contacted biological samples, especially those that may contain microorganisms. An autoclave cleans materials with pressurized steam at a high temperature. The steam and heat work together to kill microorganisms on the loaded items. For this reason, effective autoclaving depends on good steam flow and strong heat transfer. Packaging matters because it...

fume hoods and laminar flow cabinets

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handling air- and water-sensitive chemicals using a schlenk line

handling-air-and-water-sensitive-chemicals-using-a-schlenk-line

Schlenk Line Setup for Inert Atmospheres

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proper operation of vacuum based equipment

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Vacuum Pumps for Laboratory Procedures

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operating the glovebox

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operation of high-pressure reactor vessels

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Safe Use of Pressure Vessels in Chemistry

High-pressure reactor vessels are used in synthetic chemistry to handle gases safely during reactions. They help chemists carry out gas-based processes such as hydrogenation, oxidation, and amination. These reactions often use hydrogen, oxygen, or ammonia, which are important reagents in the lab. These gases usually dissolve poorly in standard reaction mixtures. Because of that, high pressure is needed to make the reaction proceed at a useful rate. The vessel creates a sealed space that keeps...

decontamination for laboratory biosafety

Lab Cleaning Methods for Biosafety

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proper waste disposal

Lab Waste Management and Safety

Lab waste management and safety are essential parts of any laboratory activity. Waste generated during work must be handled correctly from the moment it is created until it reaches its final offsite disposal facility. Poor waste disposal can seriously harm public health and the environment. A waste management system should be planned before any lab work begins. This system guides how waste is collected, stored, and prepared for removal. For hazardous waste, special rules apply because its...

lab safety

本系列提供了使用危险材料和设备时应遵循的安全指导原则。它涵盖了个人防护装备，电气安全和一般应急指南等普遍主题，以及化学和生物实验室的一些特定安全程序。

Learn how autoclaves decontaminate lab materials with pressurized steam, why steam flow and heat transfer matter, and what safety steps to follow.

Autoclaving Lab Materials Safely | JoVE K12

Chemistry

CHEM.1

CHEM.1.C

Chemistry and Physics

IPC.1

IPC.1.C

Robert Rioux

Overview

Principles of Autoclave Use

Preparing Samples for the Autoclave

Using the Autoclave

Summary

The Scientific Method

The scientific method is a detailed, empirical problem-solving process used by biologists and other scientists. This iterative approach involves ...

Inductive Reasoning

Inductive reasoning is a form of logical thinking that uses related observations to arrive at a general conclusion. It is uncertain and operates in ...

Deductive Reasoning

Deductive reasoning, or deduction, is the type of logic used in hypothesis-based science. In deductive reasoning, the pattern of thinking moves in the ...

Correlation and Causation

Correlation and CausationStatistical tests can calculate whether there is a relationship, or correlation, between independent and dependent variables. A ...

Ask questions and define problems related to specialized topics of study based on observations or information from text, phenomena, models, or investigations;

112.51.c.1.A.

Scientific Method

The scientific method is used to solve problems and explain phenomena. The development of the scientific method coincided with changes in philosophy ...

Demonstrating the Scientific Method with Tree Branches
ExpandObtain branches containing a few dozen typical leaves from five different local tree species. ...

Observation and Background Research
ExpandNOTE: The first two steps in the scientific process involve studying and observing natural phenomena of ...

Problem-Solving

A problem-solving strategy is a plan of action used to find a solution. It involves trial and error, algorithms, heuristics, and insights. Effective ...

Problem Definition

Problem definition is the initial and the most essential step in research.
The researcher identifies and explains the specific issue or gap to be ...

Apply science practices related to specialized topics of study to plan and conduct investigations or use engineering practices to design solutions to problems;

112.51.c.1.B.

Proper Personal Protective Equipment

Robert M. Rioux &#38; William A Elliott, Pennsylvania State University, University Park, PA
Hazards are many and varied in the laboratory, but the right ...

Emergency Eyewash and Shower Stations

Robert M. Rioux, Pennsylvania State University, University Park, PA
The Occupational Safety and Health Administration (OSHA) mandates that an emergency ...

Electrical Safety

Robert M. Rioux &amp; Suprita Jharimune, Pennsylvania State University, University Park, PA
Among the many hazards present in the laboratory, electrical ...

Working with Centrifuges

Source: Robert M. Rioux &amp; Suprita Jharimune, Pennsylvania State University, University Park, PA
Earth's gravitational force is capable of separating ...

Working with Hot and Cold Sources

Source: Robert M. Rioux &amp; Suprita Jharimune, Pennsylvania State University, University Park, PA
Working with extreme temperatures, both high and low, ...

Guidelines in Case of a Laboratory Emergency

Robert M. Rioux &amp; Zhifeng Chen, Pennsylvania State University, University Park, PA
The most common laboratory emergencies include chemical spills, ...

Chemical Storage: Categories, Hazards And Compatibilities

Source: Robert M. Rioux &amp; Taslima A. Zaman, Pennsylvania State University, University Park, PA
While the use of various chemicals in experimental ...

Safe Handling of Mineral Acids

Source: Robert M. Rioux &#38; Taslima A. Zaman, Pennsylvania State University, University Park, PA
A mineral acid (or inorganic acid) is defined as a ...

Handling Chemical Spills

Source: Robert M. Rioux &#38; Taslima Zaman, Pennsylvania State University, University Park, PA
Since chemicals are commonly used for laboratory research ...

Fume Hoods and Laminar Flow Cabinets

Robert M. Rioux &amp; William A. Elliott, Pennsylvania State University, University Park, PA
Fume hoods and laminar flow cabinets are engineering controls ...

Decontamination for Laboratory Biosafety

Robert M. Rioux and Zhifeng Chen, Pennsylvania State University, University Park, PA
Decontamination is essential for laboratory biosafety, as the ...

Proper Waste Disposal

Robert M. Rioux and Taslima A. Zaman, Pennsylvania State University, Pennsylvania, PA

Users are responsible for the proper disposal of the waste ...

Use appropriate safety equipment and practices during laboratory, classroom, and field investigations as outlined in Texas Education Agency-approved safety standards;

Common Lab Glassware and Uses

Source: Laboratory of Dr. Neal Abrams &#8212; SUNY College of Environmental Science and Forestry
Glassware is a regular appearance in the professional ...

Lab Techniques

Measurement Accuracy
The ability to repeat an experiment and get the same results, or reproducibility, is essential in scientific research. However, ...

Source: Smaa Koraym at Johns Hopkins University, MD, USA

	Preparation of LaboratoryExpand
	Here we show the laboratory preparation for 10 students ...

Source: Smaa Koraym at Johns Hopkins University, MD, USA

	Determining the Density of an EggExpand
	Density is a measure of how compact a substance or ...

Proper Lab Notebook Keeping

Organic Lab Notebook Keeping
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Source: Lara Al Hariri at the University of Massachusetts Amherst, MA, USA

	Preparation of the LaboratoryExpand
	Here, we show the laboratory preparation ...

Source: Lara Al Hariri and Ahmed Basabrain at the University of Massachusetts Amherst, MA, USA

	Nitration of Methyl BenzoateExpand
	From this lab onward, ...

Use tools appropriate to the specialized topic of study;

112.51.c.1.D.

SI Units: 2019 Redefinition

Measurement is integral to analytical chemistry. Each record comprises a number &#8211; denoting the magnitude&#160; &#8211; and the unit &#8211; a ...

International System of Units

The International System of Units, known as the SI system, is a universally accepted measurement system recognized and used worldwide. The SI system is ...

Measurement: Standard Units

Every measurement provides three kinds of information: the size or magnitude of the measurement (a number), a standard of comparison for the measurement ...

Measurement: Derived Units

The International System of Units or SI system, by international agreement, has fixed measurement units for seven fundamental properties: length, mass, ...

Collect quantitative data using the International System of Units (SI) or qualitative data as evidence as appropriate to the specialized topic of study;

112.51.c.1.E.

Review and Preview

Data are individual items of information obtained from a population or sample. Data may be classified as qualitative (categorical), quantitative ...

What is a Frequency Distribution

A frequency is the number of times a value of the data occurs. The sum of all the frequency values represents the total number of students included in the ...

Cumulative Frequency Distribution

A cumulative frequency distribution is another type of frequency distribution. Instead of reporting how many data values fall in some classes, it reports ...

Ogive Graph

An ogive graph is sometimes called a cumulative frequency polygon. It is one type of frequency polygon that shows cumulative frequency. In other words, ...

Histogram

The histogram is a graphical representation in the x-y form of data distribution in a data set. The horizontal x-axis is labeled with what the data ...

Relative Frequency Histogram

The relative frequency depicts the proportion of data points that have each value. The frequency tells the number of data points that have each value. ...

Scatter Plot

The most common and easiest way to display the relationship between two variables, x and y, is a scatter plot. A scatter plot shows the direction of a ...

Time-Series Graph

A time-series graph is a line graph with repeated measurements taken at successive intervals of time. It is also called a time series chart. To construct ...

Bar Graph

A bar graph is also called a bar chart and consists of bars that are separated from each other. It either uses horizontal or vertical bars to show ...

Multiple Bar Graph

As the name suggests, a multiple bar graph is the same as a bar graph but has multiple bars to depict relationships between different data values. One can ...

Pareto Chart

A Pareto chart is a bar graph or a combination of both line and bar graphs. The bar lengths represent the individual values or the frequency, while the ...

Pie Chart

A pie chart (or a pie graph) is a circular graphical chart or a pictorial representation of categorical data. It is divided into slices of pie each ...

Organize quantitative or qualitative data using representations appropriate to the specialized topic of study such as scatter plots, line graphs, bar graphs, charts, data tables, diagrams, scientific drawings, and student-prepared models;

112.51.c.1.F.

Modeling and Similitude

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This setup predicts the ...

Typical Model Studies

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...

design-example-creating-a-hydraulic-model-of-a-dam-spillway

Design Example: Creating a Hydraulic Model of a Dam Spillway

A hydraulic model of a dam spillway is often created by scaling down the design, typically to a 1:15 ratio, to replicate key flow characteristics such as ...

Models, Theories, and Laws

Scientists frequently use models to help them comprehend a specific collection of phenomena. In physics, a model is a condensed version of a physical ...

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Concepts and Prototypes

The human nervous system translates sensory stimuli into neural impulses, which the brain processes into information as thoughts and memories, forming ...

Molecular Models

Physical models representing molecular architectures of chemical compounds play essential roles in understanding chemistry. The use of molecular models ...

Develop and use models to represent phenomena, systems, processes, or solutions to problems as appropriate to the specialized topic of study; and

112.51.c.1.G.

Scientific and engineering practices. The student, for at least 40% of instructional time, asks questions, identifies problems, and plans and safely conducts classroom, laboratory, and field investigations to explain phenomena or design solutions using appropriate tools and models. The student is expected to:

Autoclaving Lab Materials Safely

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