DOCSLIB.ORG

The Autoclave Safety Poster

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
The Autoclave Safety Poster Procedures for Using Steam Autoclaves QUESTIONS: Contact DRS, 217-333-2755, for additional information. CHOOSE PROPER OPERATING CONDITIONS: MAINTENANCE & CARE: DECONTAMINATING BIOHAZARDOUS Check the Pressure: • Sterilization will occur only when the conditions of time, temperature, pressure, and humidity have been met. WASTE BAGS: Check all pressure gauges. • Incorrect selection of time or exhaust cycle may damage the autoclave, cause liquid to boil over, or bottles to break. • Use autoclavable, high strength Jacket pressure gauge should be a • Proper use of autoclave will minimize the chance of serious injury. polymer bags imprinted with minimum of 15 psi (refer to Biological Waste Liquids Dry Items Glassware chemical indicator (if possible). Items manufacturers’ instructions for (Gravity Cycle) (Liquid Cycle) (Gravity Cycle) (Gravity Cycle) • Add approximately 250 mL water to maximum psi). Loosen caps or use a Dirty: Place in middle of bag before closing and transporting Open the bag >2", Fabrics Wrap; vented closure, the pan; to autoclave area. Use Sterilization Indicators Preparation Place in tray, Instruments: Clean, Fill containers no more Clean: wash, rinse, Place indicator if needed dry, lay in pan • Place bag in a polypropylene or Visual indicators, e.g., than 75% capacity wrap stainless tray before autoclaving. chemical/tape indicators, measure Dirty: In detergent and Fabrics: Separated, on pan; • Open the bag at least 2“ to allow one or more physical conditions of Placement in Autoclave In the center Upright in pan edge; Clean: On side or the autoclave cycle. Instruments: Flat steam to enter. inverted Mechanical indicators record the • When complete, the sterilization time-temperature profile attained indicators must show adequate Temperature 121oC 121oC 121oC 121oC during a cycle. decontamination before disposal as Biological indicators, i.e., spore municipal solid waste. 22 min. for volumes 60-120 min. • Repeat cycle if sterilization vials, are the only approved way to Treatment Time in <100mL; depending on load size 30-60 min. 30-60 min. Minutes 40 min. for volumes indicators do not show evidence of prove sterility. and packing density >100mL sterilization. NOTIFY FACILITIES SUPPORT • Overbag biohazardous waste bag Dirty: Slow exhaust; Exhaust Cycle Slow exhaust Slow exhaust Fast exhaust and dry with an opaque trash bag before WHEN A PROBLEM OCCURS: Clean: Fast/dry No steam. placing in the regular trash. Valves leaking. Avoid puncturing bags. Check reference for Glassware with cracks or Hot bottles may explode. Door gasket deterioration. Notes: Overbag and dispose of proper packaging deep scratches may Let cool before moving. Reminders properly. methods crack Jacket pressure below 15 psi. • Store biohazardous waste in a Temperature not reaching 121°C CAUTION: closed leak-proof container. (250°F). • NEVER AUTOCLAVE FLAMMABLE, REACTIVE, CORROSIVE, TOXIC or RADIOACTIVE MATERIALS, e.g., bleach. • Never allow waste to accumulate in Erratic temperature or pressure • Materials that melt (plastic lab wear) at ≥ 121°C will block chamber exhaust drain if not placed in a shallow the lab. during cycle. autoclave pan able to withstand that temperature. • Never leave waste unattended. Steam escapes from around door • Use caution when increasing autoclave temperature to 135oC because plastics (including some plastic pans) • Clean up leaks and spills with a during cycle. melt at this temperature, causing difficult clean-up and damage to temperature sensors. suitable disinfectant. Water on floor or in chamber after • Always wear safety glasses, goggles, or face shield, lab coat or apron, and heat-protective non-asbestos a cycle. gloves when opening door or removing item(s) from autoclave. Articles very wet after a fast • Do not mix loads that require different exposure times and exhaust. Reference: “Using the Gravity exhaust or dry cycle. • Open door only after chamber pressure returns to zero. Leave door open for several minutes to allow Displacement Steam Autoclave in the Excessive steam in area during pressure to equalize and for materials to cool. Biomedical Laboratory” cycle or after opening door. • Open door slowly. Beware of rush of steam or water. DHHS/PHS/HIH/DS .
Load more

Recommended publications
  • Autoclave Quick Guide
    Office of Biological Safety Autoclave Operation Quick Reference Guide Training: ALL users MUST undergo documented training for operation of the autoclave. Record each use of the autoclave in a log: Date, User ID, cycle type and nature of material in load Personal Protective Equipment (PPE) needed: ¾ Wear eye protection, lab coat, gloves along with heat resistant gloves. Rubberized apron, sleeve guards and face shield are recommended when autoclave is hot or splash risk is present. Hints and Precautions ¾ Become familiar with the manufacturer’s operations manual of your autoclave model(s). ¾ Plastics used for autoclaving MUST be labeled as autoclavable – otherwise the plastic will melt. ¾ Waste bags to be autoclaved must be loosely packed and not more than 2/3 filled. Steam must be able to penetrate to all contents of the bag. ¾ Sharps or pointed hard objects should not be placed directly into an autoclave bag; a thicker or rigid container must be used (such as a sharps container). ¾ Avoid overfilling an autoclave with loads or allowing a load to contact the chamber walls. ¾ Transferring waste contents from an overfilled bag to another bag should be avoided! This practice can lead to injury and/or exposure to contaminants. ¾ Do not leave an autoclave operating unattended for long periods of time; operation should be monitored periodically during a cycle in case of failure. ¾ Never autoclave solvents, combustible, volatile, flammable, radioactive or corrosive materials (e.g. ethanol, methanol, acids, bases, phenol) ¾ Remove extraneous items and combustible materials from around the autoclave exterior. Loading ¾ Follow manufacturer’s loading instructions for your autoclave model ¾ Transport loads on a cart and in secondary containers to reduce spills ¾ Clean item/container sterilization Loosen caps or lids to avoid dangerous pressure build-up during cycle Place containers in a tray and load the tray into the autoclave – this is easier to load and unload and will catch spills.
    [Show full text]
  • Pouring Plates from Prepared Bottled Media
    Pouring Plates from Prepared Bottled Media Primary Hazard Warning Never purchase living specimens without having a disposition strategy in place. When pouring bottles, agar is HOT! Burning can occur. Always handle hot agar bottles with heat-protective gloves. For added protection wear latex or nitrile gloves when working with bacteria, and always wash hands before and after with hot water and soap. Availability Agar is available for purchase year round. Information • Storage: Bottled agar can be stored at room temperature for about six months unless otherwise specified. Never put agar in the freezer. It will cause the agar to breakdown and become unusable. To prevent contamination keep all bottles and Petri dishes sealed until ready to use. • Pouring Plates • Materials Needed: • Draft-free enclosure or Laminar flow hood • 70% isopropyl alcohol • Petri dishes • Microwave or hot water bath or autoclave 1. Melt the agar using one of the following methods: a) Autoclave: Loosen the cap on the agar bottle and autoclave the bottle at 15 psi for five minutes. While wearing heat-protective gloves, carefully remove the hot bottle and let it cool to between 75–55°C before pouring. This takes approximately 15 minutes. b)Water Bath: Loosen the cap on the agar bottle and place it into a water bath. Water temperature should remain at around 100°C. Leave it in the water bath until the agar is completely melted. While wearing heat- protective gloves, carefully remove the hot bottle and let it cool to between 75–55°C before pouring. c) Microwave: Loosen the cap on the agar bottle before microwaving.
    [Show full text]
  • Biohazardous Waste Handling for Eastern Kentucky University May 2018
    Biohazardous Waste Handling For Eastern Kentucky University May 2018 Approved by The University Laboratory Safety Biohazard Subcommittee 1 Table of Contents Page Introduction 3-4 Definitions 5-6 Responsibilities and roles 7-9 Main body of document 10-17 Appendix 1 18-22 Autoclave procedures in SB 18-20 Autoclave procedures in Disney 218 21-22 Appendix 2 copy of IACUC form H: Use of hazardous agents 23 2 3 Introduction This document has been prepared to provide guidance to Eastern Kentucky University employees and students in the use and disposal of biohazardous materials in compliance with regulatory requirements. For this document “biohazardous waste” is defined as any discarded material which might include infectious laboratory materials or agents regulated by federal, state, and local authorities. At a minimum, the following categories should be considered as biohazardous waste materials: 1. Cultures and stocks of infectious biological agents, including laboratory waste, discarded live or attenuated viruses or related agents, culture dishes and other laboratory supplies used in the production or use of these agents, and any other related devices. 2. Human blood is a biohazardous waste. Human blood should treated in accordance with the “Eastern Kentucky University Bloodborne Pathogens Exposure Control Plan”. This plan can be found at the Risk Management Insurance/ Environmental Health and Safety website https://ehsrmi.eku.edu/occupational-safety. 3. Sharps: These are defined as needles, syringes, scalpels, etc., as well as any object sharp enough to puncture the skin (i.e. microscope slides, cover slips) that is used in the laboratories that could possibly come in contact with material that may be considered biohazardous waste.
    [Show full text]
  • The Eppendorf Mechanical Pipettes Research® Plus and Reference 2 – Fully Autoclavable, Easy Adjustment, Quick and Simple Maintenance
    APPLICATION NOTE No. 198 The Eppendorf Mechanical Pipettes Research® plus and Reference 2 – Fully Autoclavable, Easy Adjustment, Quick and Simple Maintenance Kornelia Ewald, Ulrike Gast, Eppendorf AG, Hamburg, Germany Abstract Research plus and Reference 2 pipettes are easy to clean safe to perform. This Application Note demonstrates the and maintain. Hence, routine maintenance can be easily special features of maintaining and cleaning Research performed by the user, saving valuable time. Readjust- plus pipettes and provides detailed information on ment of the pipette for liquids whose physical properties changing the pipette‘s adjustment. diff er signifi cantly from those of water is also easy and Introduction Research plus and Reference 2 are manual pipettes with Adjustment for specifi c liquids or altitude diff erent features and functional principles (e.g. one-button / During production, piston stroke pipettes are adjusted two-button operation). Fulfi lling the highest requirements, to distilled water under certifi ed measuring conditions. both pipettes have equal functionalities in terms of decon- To indicate the adjustment, all Research plus pipettes carry tamination, maintenance and adjustment. In order to meet an adjustment seal (Fig. 1). If necessary, adjustment for the high requirements of a modern laboratory, high quality specifi c liquids or for altitude can be carried out easily by piston stroke pipettes should be partly or fully autoclavable the user. The red adjustment seal, which is applied to the as well as UV resistant. adjustment opening following an adjustment, serves to The Eppendorf Research plus and Reference 2 pipettes can visualize a change of adjustment settings (Fig. 1). be decontaminated either by UV light or by autoclaving the entire instrument.
    [Show full text]
  • Elara11 Autoclave Quick Installation Guide
    OVER YEARS S ince 1925 Elara11 Autoclave Quick Installation Guide 1. Examine the outer carton and autoclave for any signs of damage. Immediately notify your dealer or Tuttnauer USA of any signs of damage. 2. To avoid injuries, lifting and carrying should be done by two people. • Lifting straps have been installed for your convenience. Lifting straps are for one time use only and should be removed and discarded after initial set up. 3. Place the sterilizer on a rigid level surface. The counter top or stand must be able to support 275lbs lbs Minimum 24”depth 4. The minimum depth of the counter top needs to be 24 inches. counter top required 5. It is mandatory to leave a minimum of 2” clearance between the back of the Elara11 and the wall. If located in a cabinet, the rear panel of the cabinet must be removed for proper air circulation. Failure to provide the needed clearance will result in failed cycles. 6. Side clearances should be a minimum of 2”. 7. Make sure all the feet are on the autoclave and none of them have been lost. 8. Connect the power cord to the socket on the rear of the autoclave; then plug it into the supply outlet. Power switch a. This unit requires a 230 volt 1 or 2 phase 15A supply. b. The acceptable operating voltage range is 220 to 235 volts. c. The installation of a Buck/Boost transformer (0.5KVA) may be required to meet the acceptable operating voltage. d. The supply outlet must be a properly grounded outlet.
    [Show full text]
  • Cleaning and Decontamination Guide for Sartorius Pipettes
    Operating Manual Cleaning and Decontamination Guide for Sartorius Pipettes Version 1 / 2018 / 11 Contents Contents 1 Cleaning and Decontamination in Pipette Maintenance . 4 2 Cleaning Guide . 5 2 .1 Cleaning the outer surface of the pipette (daily) . 5 2 .2 Cleaning the lower part of the pipette (every three months) . 5 3 Application-Specific Cleaning and Decontamination of Pipettes . 10 3 .1 When cleaning Sartorius pipettes for specific applications . 10 3 .2 Cleaning and decontamination reagent quick-guide . 11 3 .3 Cleaning agents and their activities . 11 3 .4 Cleaning procedures . 12 3 .5 Decontamination agents and their activities . 12 3 .6 Decontamination procedures . 13 4 Autoclaving Guide . 14 4 .1 Mechanical pipettes . 14 4 .2 Electronic pipettes . 14 4 .3 Autoclaving quick-guide . 15 4 .4 Drying of pipettes . 15 5 Summary of Pipette Parts . 16 5 .1 Mechanical pipettes parts and materials . 16 5 .1 .1 Tacta® mechanical pipette . 16 5 .1 .2 mLine® mechanical pipette . 17 5 .1 .3 Proline® Plus mechanical pipette . 17 5 .1 .4 Proline® mechanical pipette . 18 5 .2 Electronic pipette parts and materials: Picus and Picus® NxT . 18 6 Contact . 19 6 .1 Further information about pipette cleaning, decontamination and maintenance . 19 6 .2 Pipette maintenance, servicing or calibration . 19 Cleaning and Decontamination Guide for Sartorius Pipettes 3 Cleaning and Decontamination in Pipette Maintenance 1 Cleaning and Decontamination in Pipette Maintenance Pipettes are precision instruments whose performance can be significantly impacted if not adequately maintained . To maintain the level of purity that is critical in many laboratory applications, adequate cleaning and decontamination of pipettes is necessary .
    [Show full text]
  • The Effect of Autoclave Resterilisation on Polyester Vascular Grafts
    CORE Metadata, citation and similar papers at core.ac.uk Provided by Elsevier - Publisher Connector Eur J Vasc Endovasc Surg 18, 386±390 (1999) Article No. ejvs.1999.0891 The Effect of Autoclave Resterilisation on Polyester Vascular Grafts G. Riepe∗1, M. S. Whiteley2, A. Wente1, A. Rogge1, A. SchroÈ der1, R. B. Galland2 and H. Imig1 1Department of General, Vascular and Thoracic Surgery, General Hospital of Hamburg±Harburg, Germany; 2Department of Surgery, Royal Berkshire Hospital, Reading, U.K. Objectives: polyester grafts are expensive, single-use items. Some manufacturers of uncoated, woven grafts include instructions for autoclave resterilisation to be performed at the surgeon's own request. Others warn against such manipulation. Theoretically, the glass transition point of polyester at 70±80°C and the possible acceleration of hydrolysis suggest that autoclave resterilisation at 135°C might be a problem. Materials and methods: a DeBakey Soft Woven Dacron Vascular Prosthesis (Bard) and a Woven Double Velour Dacron Graft (Meadox) were autoclave-resterilised 0 to 20 times, having been weighed before and after sterilisation. Tactile testing was performed. Mechanical properties were examined by probe puncture and single-®lament testing, the surface was examined by scanning electron microscopy and the degree of hydrolysis by infra-red spectroscopy. Results: tactile testing revealed a change of feeling with increasing cycles of resterilisation. Investigation of weight, textile strength, single-®lament strength, electron microscopy of the surface and infra-red spectroscopy showed no change of the material. Conclusions: changes felt are presumably a surface phenomenon, not measurably affecting strength or chemistry of material after autoclave resterilisation.
    [Show full text]
  • Transforming Plasmid DNA Into Electrocompetent Cells
    Transforming plasmid DNA into electrocompetent cells 1. Clean and dry electroporation cuvettes throroughly on the cuvette washer. Chill on ice and allow to air dry. Use one cuvette for each DNA sample you are transforming. 2. Dialyze your DNA sample(s) using a nitrocellulose filter and DI water. • Fill a Petri dish with DI water. • Place a single nitrocellulose filter paper on the surface of the water – shiny side up. If you are dialyzing more than one sample, it helps to cut a small notch in the paper to serve as a “key”. This helps to identify/keep track of your samples. • Spot DNA samples onto the surface of the paper. Space out the samples a little bit because they will expand in volume as they dialyze. • Dialyze your sample for 10-15 minutes. *This step is particularly important if the DNA has undergone any previous manipulations that have introduced salts (adding buffers for ligation, restriction enzyme digestions, etc.). Too much salt in the DNA will cause your sample to arc when electroporating. This will kill the cells and significantly reduce the efficiency of transformation. If your sample arcs, it may still be worth while to plate your cells. However, it is likely that you will get very few (if any) colonies. You should repeat this sample. Main factors that can cause arcing - too much salt in your DNA, water on the outside of the cuvette, oil on the outside of the cuvette from handling it too much without gloves, too much salt in the cells. 3. Thaw electrocompetent cells on ice.
    [Show full text]
  • Egyptians Used Fire for Sterilisation • Greeks Burned Sulphur for Fumigation • Advanced Techniques
    • Egyptians used fire for sterilisation • Greeks burned sulphur for fumigation • Advanced techniques To Prevent • Transmission of diseases. • Spoilage of food. • Contamination of pure cultures in laboratories. • Interference of unwanted microbes in industrial process. • Research studies. Choice of Anti microbial agents depends on • Type of microbe • Stages of growth and number • Surroundings 5 METHODS/AGENTS OF STERILIZATION Classification of Sterilization: Sterilization Physical methods. Chemical methods. 6 Physical agents: Sunlight Drying Heat Dry heat: flaming, incineration, hot air Moist heat: pasteurization, boiling, steam under pressure. Filtration: candles, asbestos pads, membranes Radiation Ultrasonic vibrations. Sun light: • Sun light: – Active germicidal effect due to its content of ultraviolet rays . – Natural method of sterilisation of water in tanks, rivers and lakes. 12/2/2012 Dr.T.V.Rao MD 7 Drying: Moisture is essential for growth of bacteria. Drying in air has deleterious effect on many bacteria. However, spores are unaffected. Heat: • Most reliable method of sterilization and should be the method of choice. Dry Heat & Moist Heat. The factors influencing sterilization by heat: • Nature of heat-dry or moist • Temperature and time • Number of microorganisms present • Characteristics of organisms –species, strain, sporing capacity • Type of material from which organism have to be eliminated. • Killing effect is due to protein denaturation, oxidative damage and toxic effect of elevated level of electrolytes. • Killing effect of moist heat due to denaturation and coagulation of proteins. Thermal Death Time:TDT “Minimum time required to kill a suspension of organisms at a predetermined temperature in a specified environment. Thermal death time is inversely proportional to temperature. TDT is increased in presence of organic substance, proteins, nucleic acid, starch, gelatin , sugar , fats, oils.” Heat : • Dry heat: • 1.Red heat 2.Flaming 3.Incineration 4.Hot air oven 12/2/2012 Dr.T.V.Rao MD 11 Dry heat: 1.
    [Show full text]
  • Autoclave Safety Purpose
    Autoclave Safety Purpose: Sterilization refers to the complete killing of all living organisms, including spores. Common sterilization techniques include the application of wet heat, dry heat, chemicals, and radiation. The type of material, the container, and quantity of items to be sterilized determines which method to use. Various pieces of equipment are used for sterilization in laboratory animal facilities, but the autoclave is the primary means of sterilizing materials. Despite built-in safeguards, an autoclave presents the possibility of serious injury to personnel’s from hot surfaces and from the release of steam. It is important, therefore, that laboratory personnel understand the proper operation, limitations, and safeguards for sterilization by autoclaving. ASSOCIATED RISKS: Autoclaves are sterilizers using high pressure and high temperature steam. The potential safety risks for the operators are: • Heat burns -from hot materials and autoclave chamber walls and door • Steam burns -from residual steam coming out from autoclave and materials on completion of cycle • Hot fluid scalds- from boiling liquids and spillage in autoclave. • Hand and arm injuries when closing the door. • Body injury if there is an explosion Autoclaves are used in laboratories to sterilize equipment, instruments, and infectious waste. No one who has not received training in autoclave procedure or is not working under the supervision of an experienced autoclave worker should attempt to operate the autoclave. In addition, users should read and understand the owner’s manual from the particular model of autoclave that they are using. SAFETY PRECAUTIONS: • All operators must receive training on the safe operation of the autoclave prior to using the equipment.
    [Show full text]
  • 3M™ Petrifilm™ Aerobic Count Plates Introduction SCIENTIFIC Petrifilm™ Plates Are Designed to Determine Total Aerobic Bacterial Populations
    3M™ Petrifilm™ Aerobic Count Plates Introduction SCIENTIFIC Petrifilm™ plates are designed to determine total aerobic bacterial populations. The easy-to-use BIO FAX! Petrifilm plates contain a film coated with nutrients and gelling agents. You will be able to monitor environmental bacterial counts using a variety of methods. Three environmental sampling methods are described as well as the standard inoculation procedure. Concepts • Microbiology • Plate counting Materials Petrifilm Aerobic Count Plates Incubator Petrifilm Spreader Pipet Chip clips Tape, double-sided Cotton swab, sterile Test tube, screwtop Safety Precautions Wear chemical-resistant gloves while working with microbial organisms. After use, Petrifilm Aerobic Count Plates will contain viable cultures. Do NOT reopen the Petrifilm plates or handle unnecessarily. Wash hands thoroughly with soap and water before leaving the laboratory. Extreme caution should be exercised when handling culture materials, and students should be trained in aseptic techniques. Always clean work areas and wash hands after working with microbiological materials. Procedures Standard Inoculation 1. Place the Petrifilm Aerobic Count Plate on a flat surface. Carefully peel open the Petrifilm plate being careful not to touch the nutrient gel with your fingers. (See Figure 1.) 2. With a pipet perpendicular to the Petrifilm plate, place 1 mL of sample (inoculum) onto the center of the bottom film. (See Figure 2.) 3. Release the top film; allow it to drop. Do not roll top film down. (See Figure 3.) Figure 1. Figure 2. Figure 3. FLAT side RIDGE side Figure 4. Figure 5. Figure 6. © 2016 Flinn Scientific, Inc. All Rights Reserved. Publication No. 10163 1 061616 3M™ Petrifilm™ Aerobic Count Plates continued 4.
    [Show full text]
  • Procedures for Decontamination by Autoclaving
    Procedures for Decontamination by Autoclaving Purpose: Biohazardous waste material and sharps containers generated within research and teaching facilities are required to be decontaminated in laboratory (or departmental) autoclaves and disposed of using the appropriate waste streams. The procedures below serve as guidelines to help autoclave users ensure safe and effective processing. 1. Select appropriate containers or bags for 5 POLYPROPYLENE AUTOCLAVE BAG collecting materials to be autoclaved. For biohazardous dry solid materials a. Collect in polypropylene AUTOCLAVE bags: BSL-1 waste Æ Clear bags, no symbol BSL-2 waste Æ Orange bags, symbol BSL-3 waste Æ Red bags, symbol b. DO NOT use the red bags that come with the Regulated Medical Waste (RMW) boxes for 9 SHARPS PENETRATING BAG initial waste collection. They are not meant to be autoclaved. c. Ensure that bags are free of sharp objects that may puncture bags. Autoclave bags are tear resistant, but can be punctured or burst in the autoclave. d. Fill bags only 2/3 full. e. Ensure adequate steam penetration by creating an opening of at least one inch in the 5 CLOSURE 9 CLOSURE bag’s closed top. f. On autoclaves which have no Prevacuum cycle, water can be carefully added to bags of waste run on Solids/Gravity cycle if needed to achieve effective decontamination. (Steam created inside the bag during processing aids in reaching appropriate temperature.) 9 OVERFILLED For biohazardous sharps: a. Collect in commercially available Sharps containers with lids or closures. Containers must not be tightly sealed shut AND MUST NOT BE OVERFILLED. For biohazardous liquids: 5 PROPER CLOSURES a.
    [Show full text]