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. Indossare adeguati dispositivi di protezione individuale (DPI)
Le condizioni operative ad alta pressione e temperatura incontrate durante l'autoclave comportano potenziali rischi di bruciatura o esplosione di calore. I DPI appropriati includono un cappotto da laboratorio, occhiali di sicurezza, guanti resistenti al calore e scarpe chiuse; questi devono essere indossati durante il carico e lo scarico dei materiali dall'autoclave. Quando si tratta di campioni liquidi, indossare un grembiule e una visiera come DPI aggiuntivi.
2. Preparazione dei materiali da caricare
<ol><li>Identificare il materiale compatibile con l'autoclave e il materiale incompatibile con l'autoclave Sebbene l'autoclave rappresenti un metodo comunemente usato ed economico per la decontaminazione, non tutti i materiali sono adatti per l'autoclave a causa delle condizioni di lavoro ad alta pressione e temperatura. In generale, tutti i materiali che evaporano o si fondono sotto il vapore ad alta pressione non sono compatibili con l'autoclave. Ciò include materiali come solventi organici, plastica scarsamente resistente al calore come polistirene, polietilene e metalli escluso l'acciaio inossidabile. Mai autoclave infiammabile, corrosiva, tossica, reattiva o radioattiva. I materiali compatibili per l'autoclave includono vetro borosilicato Pyrex o tipo I, polipropilene, policarbonato, guanti, acciaio inossidabile, punte per pipette, carta (inserita in sacchetti per autoclave) e soluzioni multimediali.</li>
	<li>Materiale di imballaggio Dopo aver identificato i materiali compatibili con l'autoclave, è importante anche un imballaggio adeguato poiché garantisce sicurezza e autoclave efficiente. Ecco i principi di base quando si imballano i materiali:<ol>
<li>Utilizzare sempre un contenitore secondario in polipropilene o acciaio inossidabile.</li>
		<li>Per i campioni liquidi, riempire il flaconcino di circa 1/2 pieno e allentare i tappi o utilizzare chiusure ventilate. Non riempire mai i contenitori oltre i 2/3 pieni con il tappo completamente serrato.</li>
		<li>Ispezionare il recipiente di vetro per assicurarsi che non ci siano crepe. Se vengono identificate crepe, smaltire i bicchieri in un recipiente appropriato. Mai autoclave un noto vaso di vetro rotto.</li>
		<li>Utilizzare sempre sacchetti compatibili con l'autoclave per confezionare i rifiuti. I sacchetti devono avere nastri di indicazione su di esso e devono essere aperti alle estremità prima del caricamento per consentire il vapore all'interno del sacchetto.</li>
		<li>Non sovraccaricare mai i sacchetti o il contenitore per consentire un flusso di vapore sufficiente per una sterilizzazione completa. Lascia spazio tra gli elementi.</li>
		<li>Non mescolare materiali incompatibili.</li>
		<li>Non mettere mai oggetti taglienti nel sacchetto dei rifiuti.</li>
	</ol>
</li>
</ol>3. Carica materiale
<ol><li>Controllare l'interno dell'autoclave per assicurarsi che non rimanga alcun potenziale pericolo da parte dell'utente precedente.</li>
	<li>Pulire lo scarico prima di caricare i materiali per consentire una buona circolazione.</li>
	<li>Assicurarsi che i sacchetti non tocchino le pareti interne dell'autoclave per evitare di sciogliersi.</li>
	<li>I liquidi e i materiali secchi richiedono cicli diversi e devono essere autoclavati separatamente.</li>
	<li>Chiudi saldamente la porta.</li>
</ol>4. Funzionamento dell'autoclave
<ol><li>Solo il personale addestrato dovrebbe essere autorizzato a utilizzare autoclavi. Fare riferimento al manuale dell'apparecchiatura, se necessario, poiché i protocolli operativi variano tra le diverse autoclavi.</li>
	<li>Dopo essersi assicurati che la porta dell'autoclave sia ben chiusa, impostare la temperatura dell'autoclave a 121 °C e mantenere una pressione di almeno 15 psi.</li>
	<li>Per impostare il tempo e il ciclo dell'autoclave, è necessario considerare i seguenti fattori:<ol>
<li>Raccomandazioni sull'attrezzatura dal manuale del produttore.</li>
		<li>Il tipo di materiali caricati all'interno: secco o liquido.</li>
		<li>Le quantità di materiale o di materiale caricato.</li>
		<li>Forma e dimensione del contenitore utilizzato.</li>
		<li>Conducibilità termica del contenitore e dei materiali.</li>
		<li>Scopo della decontaminazione.</li>
	</ol>
</li>
</ol>5. Scarico di materiale/i autoclavato(i)
<ol><li>Indossare DPI adeguati come descritto sopra.</li>
	<li>Assicurarsi che la temperatura e la pressione dell'autoclave siano tornate a un intervallo di sicurezza.</li>
	<li>Stare dietro la porta e aprire con attenzione la porta per rilasciare il vapore residuo all'interno dell'autoclave nella stanza.</li>
	<li>Lasciare riposare i materiali all'interno dell'autoclave per 10 minuti per rilasciare vapore o aria calda intrappolati all'interno del materiale.</li>
	<li>Non agitare alcun materiale liquido mentre lo si rimuove dall'autoclave. Se necessario, etichettare il materiale come caldo per evitare ustioni da calore a un individuo che potrebbe essere potenzialmente inconsapevole del fatto che il materiale è stato recentemente autoclavato.</li>
	<li>Attendere che il materiale si raffreddi a temperatura ambiente prima di trasportare all'interno del laboratorio o all'esterno del laboratorio. In entrambi i casi, assicurarsi di utilizzare un contenimento appropriato per il trasporto.</li>
	<li>Registrare i dettagli dei materiali e delle condizioni di autoclave nel registro di un utente.</li>
</ol>

Rispetto ai metodi di decontaminazione chimica o da radiazioni, l'autoclave rappresenta il metodo più economico e pratico in laboratorio. Tuttavia, per garantire la sicurezza (evitando la fusione, le bruciature da calore o l'esplosione), identificare materiali compatibili con l'autoclave e praticare un imballaggio, un carico e uno scarico adeguati. Garantire una sufficiente efficienza di decontaminazione richiede materiali per contenitori o sacchi e spazio sufficiente tra i materiali all'interno dell'autoclave. La manute...

uso corretto delle autoclavi

Robert M. Rioux & Zhifeng Chen, Pennsylvania State University, University Park, PA
L'autoclave è uno dei metodi più comunemente usati in laboratorio ai fini della decontaminazione. Gli articoli più comuni decontaminati con un'autoclave sono quelli che contattano campioni biologici (in genere quelli contenenti microrganismi). Un'autoclave funziona utilizzando vapore pressurizzato ad alta temperatura per uccidere i microrganismi presenti nei materiali caricati. Un flusso di vapore e un trasferimento di calore sufficienti sono essenziali per un'autoclave altamente efficiente, che sono i principi chiave da considerare quando si confezionano i materiali. La sicurezza deve essere considerata quando si lavora con un'autoclave a causa dell'alta pressione e temperatura ivi impiegate, che stabilisce anche limiti su quali materiali sono compatibili e può richiedere particolare attenzione durante l'imballaggio, il carico e lo scarico dei materiali.

Uso corretto delle autoclavi

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

L'autoclave è uno dei metodi più comunemente utilizzati in laboratorio ai fini della decontaminazione. Molti scienziati, in particolare quelli che lavorano con campioni biologici, hanno bisogno di attrezzature e terreni sterilizzati e le autoclavi forniscono un modo rapido ed economico per decontaminare i materiali. Usano il vapore ad alte temperature e pressione per uccidere i microrganismi. A causa di queste condizioni estreme, tuttavia, devono essere usati con attenzione. Questo video illustrerà come funzionano le autoclavi e come utilizzarle correttamente. In un'autoclave, i materiali vengono riscaldati con vapore a temperature di 121 gradi Celsius a pressioni di almeno 15 p.s.i. Dopo almeno 30 minuti, queste condizioni disidratano e uccidono le cellule. Le autoclavi possono sterilizzare sia materiali liquidi che solidi, ma è necessario considerare diverse impostazioni operative. I liquidi richiedono tempi più lunghi rispetto ai solidi per scaricare il vapore, al fine di prevenire l'ebollizione rapida. Inoltre, volumi maggiori di materiali liquidi impiegano più tempo a riscaldarsi fino alla temperatura corretta. Non tutti i materiali sono in grado di resistere alle temperature all'interno di un'autoclave. Pertanto, non sterilizzare mai in autoclave nulla che si sciolga, bruci, sia esplosivo o liquidi che evaporeranno. Inoltre, non sterilizzare in autoclave sostanze chimiche tossiche o radioattive. Ora vi mostreremo come preparare i campioni per l'autoclave e come utilizzarne una. Innanzitutto, seleziona i materiali che intendi sterilizzare in autoclave e utilizza solo contenitori e strumenti compatibili. Per la sterilizzazione in autoclave possono essere utilizzati pirex, vetro borosilicato, acciaio inossidabile, polipropilene e policarbonato. Sono compatibili anche altri materiali di laboratorio comuni, come puntali e scatole per pipette. La carta può essere sterilizzata in autoclave in sacchetti compatibili con l'autoclave. I rifiuti solidi possono anche essere sterilizzati in autoclave in un apposito sacchetto. Lasciare spazio in questo sacchetto per consentire al vapore di circolare e legare le estremità. Infine, prima di procedere in autoclave, assicurati di creare una piccola apertura nel sacchetto per consentire l'ingresso del vapore. Per i terreni acquosi e altri campioni liquidi compatibili, utilizzare un contenitore intatto e privo di crepe. Riempi il contenitore per circa metà e mai per più di due terzi. Per ventilare i gas da un contenitore di liquidi, allentare il tappo. Se non c'è il tappo, coprire senza stringere con un foglio di alluminio per evitare la contaminazione del materiale autoclavato. Mettere tutti i materiali in un contenitore secondario in polipropilene o acciaio inossidabile. Separare i materiali solidi e liquidi da sterilizzare in autoclave. Lascia spazio tra i materiali e non riempire eccessivamente il contenitore. Infine, attacca un pezzo di nastro adesivo per autoclave a tutti i materiali. Questo nastro è dotato di indicatori sensibili alla temperatura che diventano neri se viene raggiunta una temperatura sufficiente. Quando si utilizza un'autoclave, indossare adeguati dispositivi di protezione individuale per proteggersi dai rischi di alte temperature e pressioni. Ciò include occhiali di sicurezza DPI standard, camici da laboratorio, scarpe chiuse e guanti resistenti al calore. Utilizzare un grembiule e una visiera quando si maneggiano campioni liquidi. All'autoclave, aprire prima lo sportello e verificare la presenza di eventuali pericoli all'interno. Caricare separatamente i materiali liquidi e secchi, poiché richiedono cicli di autoclave diversi. Per evitare lo scioglimento, assicurarsi che i materiali non tocchino le pareti. Garantire un buon flusso di calore mantenendo una distanza sufficiente tra i materiali. Infine, chiudere e sigillare la porta. Autoclavi diverse hanno istruzioni operative diverse. I cicli possono essere impostati impostando manualmente la temperatura e l'ora o selezionando da un elenco preimpostato. In generale, utilizzare i cicli consigliati per il tipo di materiale che si sta decontaminando. Esistono una varietà di cicli per solidi e liquidi, nonché per diverse quantità di materiale e rifiuti da sterilizzare in autoclave. Quando si esegue un ciclo, assicurarsi che la temperatura abbia raggiunto almeno 121 gradi Celsius e la pressione di almeno 15 p.s.i.Dopo che il ciclo è terminato, assicurarsi che la pressione sia tornata a zero e che l'autoclave si sia raffreddata. Aprire con cautela lo sportello per far uscire il vapore residuo. Quindi, lasciare riposare i materiali per 10 minuti per rilasciare eventuali gas caldi. Il nastro dell'autoclave dovrebbe ora avere strisce nere o dire autoclavato su di esso, anche se questa non è una garanzia che la sterilizzazione sia avvenuta. Indossando DPI adeguati, rimuovere i materiali, facendo attenzione a non agitare i campioni di liquido. Annotare l'utilizzo nel registro, compreso il tipo di sostanze e le eventuali circostanze speciali, se si verificano. Infine, trasportare i materiali in un'area sicura o in una cappa a flusso laminare per raffreddarli. Hai appena visto l'introduzione di JoVE alle autoclavi. A questo punto è necessario capire quali materiali possono essere sterilizzati in autoclave, come prepararli e come utilizzare lo strumento in sicurezza. Grazie per l'attenzione!

Education

Didattica

JoVE Science Education

Basic Biology

Biologia di base

Lab Safety

Sicurezza in Laboratorio

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

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proper use of autoclaves

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fume hoods and laminar flow cabinets

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Questa collezione fornisce linee guida di sicurezza da seguire quando si lavora con materiali e attrezzature pericolosi. Copre argomenti universali come i DPI, la sicurezza elettrica e le linee guida generali per le emergenze, oltre ad alcune procedure di sicurezza specifiche per i laboratori di chimica e biologia.

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
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Lab Techniques

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

	Determining the Density of an EggExpand
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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

A scaled model of a dam is tested in a controlled environment to observe how it will manage water flow, turbulence, and pressure.
This setup predicts the ...

Typical Model Studies

Typical model studies analyze scaled-down systems to predict fluid behavior, focusing on interactions with free surfaces and flow around immersed bodies.
...

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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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