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MANAGEMENT AND TECHNIQUES OF SCIENCE LAB REPORT

TOOLS AND MATERIALS STERILIZATION TECNIQUES

By:
ALFATAH FATHONY

(103152440O4)

THERESIA ASMARA SEJATI

(10315244007)

AMILA RIZQI WULAN UTAMI

(10315244009)

ADITYA HILMAN PRATAMA

(10315244015)

DESTIKA SETYA PRATIWI

(10315244017)

GANIE INDRA VIANTORO

(10315244022)

NILIA FITHRIYYATI

(10315244024)

YOGYAKARTA STATE UNIVERSITY
MATHEMATHICS AND SCIENCE FACULTY
NATURAL SCIENCE STUDY PROGRAM
2011

formalin. Microorganisms are living organisms that can reproduce under the optimum . Background At this present time. Sterilization with a swab done to determine the number of microbes on the surface of the body. and rays have shorter wavelengths. Mechanically sterilization (filtration) using a porous filter is very small (micron 0:22 or 0:45 microns) so that the microbes retained on the filter. certainly in the field of research microorganisms using techniques or special ways to study microorganisms as well as to work on a laboratory scale in researching both the nature and characteristics of microorganisms. X rays. disinfectants. In principle. the use of UV light. especially experiments and observation of microorganisms. especially experiments and observation of microorganisms. and so on. This is done because the tools used in microbiology laboratories have different sterilization techniques. Objectives 1. Chemical sterilization is done by using chemicals such as alcohols. preparation and use and function of each tool and the working principles of microbiology laboratory. To know the techniques of sterilization equipment used in biological experiments. Chemicals that good is having the ability to kill microbes rapidly with low doses without damaging the material or sterilized tools. 2. The selection mechanism performed sterilization should be tailored to the nature of the material to be sterilized. Physics sterilization carried out by using the heating. Basic Theory Sterile is a term that indicates the condition with no living microorganisms. sterilization can be done in three ways: mechanically. This is done to memuddahkan berlangsungkan a study. B. we conducted this experiment to find recognition techniques. physically and chemically. To know the techniques of sterilization of materials used in biological experiments. And it is also necessary for the sterilization of knowledge about the ways or sterilization techniques. Therefore. Based on the above.TOOLS AND MATERIALS STERILIZATION TECNIQUES A. C. the course also required knowledge of the microbiology laboratory tools and techniques or how to use the tools associated with the research. The tools used in microbiology lab should also be in a state of sterile or free from germs and bacteria. viruses and fungi. Also anyway to know the techniques of sterilization of those tools.

not a single event. Because sterilization is a process. saturated steam using an autoclave. The amount and type of organic material that protects the microorganisms. wit steam sterilization. The effectiveness of any method of sterilization is also dependent upon four other factors: 1. without thorough cleaning. which removes any organic matter remaining on the instruments that could protect microorganisms during the sterilization process. dry heat (oven). Sterilization is intended to prevent contamination of equipment and materials used in microbiological testing by bacteria and viruses from the surrounding environment. 2. It is generally . The type of microorganism present. Sterilization is a process to kill any microorganisms that exist. temperature and. cracks and crevices such as the serrated jaws of tissue forceps. To be effective. and are protected by. or dry heat using an oven. sterilization requires time. Microorganisms collect in. It is much easier to kill one organism than many. even with longer sterilization times. Some microorganisms are very difficult to kill. chemical sterilants (glutaraldehydes or formaldehyde solutions) or physical agents (radiation). Sterilization should be able to kill microorganisms is the most heat resistant bacterial spores. so if grown in a medium no longer micro-organism that can multiply. surgical gloves and other items that come in direct contact with the blood stream or normally sterile tissues (Spaulding 1939). sterilization cannot be assured. contact. Finally. High-pressure steam sterilization is an effective method of sterilization but is the most difficult to do correctly (Gruendemann and Mangum 2001). Others die easily. high pressure. are the most common and readily available methods used for sterilization. 3. Blood or tissue remaining on poorly cleaned instruments acts as a shield to microorganisms during the sterilization process. Sterilization in general is a method of destruction of all forms of living organisms. all components must be carried out correctly for sterilization to occur. The number of microorganisms present.conditions for growth. 4. The number of cracks and crevices on an instrument that might harbor microorganisms. It can be achieved by high-pressure steam (autoclave). METHODS OF HEAT STERILIZATION High-pressure. Sterilization should be used for instruments. scratches.

can be used only with glass or metal objects it will melt other substances. Dry-heat sterilizers (ovens) are good in humid climates but need a continuous supply of electricity. The material used for wrapping instruments and other items must be porous enough to let steam through but tightly woven enough to protect against dust particles and microorganisms. Furthermore. Sterile instruments and other items should be used immediately unless they:  were wrapped in a double layer of muslin. the package becoming wet or anything else that will allow microorganisms to enter the package or container. especially under pressure. Certain types of contaminants. dry-heat sterilization. instruments can be sterilized in a nonelectric steam sterilizer using kerosene or other fuel as a heat source. This reemphasizes the need for thorough cleaning of objects before sterilization. STERILIZATION BY STEAM Steam is an effective sterilant for two reasons. even though the oven is operated at a much higher temperature. allowing coagulation (similar to cooking an egg white) of the sensitive inner portions of the microorganism. An event can be a tear or worn area in the wrapping. steam is an effective sterilant because any resistant. It is many times more effective in conveying this type of energy to the item than is hot (dry) air. saturated steam is an extremely effective “carrier” of thermal energy. protective outer layer of the microorganisms can be softened by the steam. especially greasy or oily materials. Wrapped sterile packs should remain sterile until some event causes the package or container to become contaminated. paper or other appropriate material prior to sterilization. potatoes can be cooked in a few minutes in a steam pressure cooker while cooking may take an hour or more in a hot-air oven. Second. . sterile container with a tight-fitting lid. while hot air does so very slowly. making them impractical in many remote (rural) areas.the method of choice for sterilizing instruments and other items used in healthcare facilities. Steam. however. or  can be stored in a dry. In a kitchen. thus hindering the process of sterilization. First. carries thermal energy to the potatoes very quickly. can protect microorganisms against the effects of steam. Where electricity is a problem. which requires use of higher temperatures.

therefore. such as Cidex. Eventually. Common disinfectants that can be used for chemical sterilization include glutaraldehydes and formaldehyde. it is also more irritating to the skin. but they are the only practical sterilants for some instruments. dry heat is a practical way to sterilize needles and other instruments. the entire object reaches the temperature needed for sterilization.STERILIZATION BY DRY HEAT When available. Also. are often in short supply and very expensive. Dry-heat sterilization is accomplished by thermal (heat) conduction. eyes and respiratory tract and is classified as a potential . because the moisture in the steam sterilization process significantly speeds up the penetration of heat and shortens the time needed to kill microorganisms. rinsing with sterile water is essential if the item must be kept sterile. but using high-pressure steam or dry-heat sterilization would damage them or equipment is not available (or operational). Both glutaraldehydes and formaldehyde require special handling and leave a residue on treated instruments. Although formaldehyde is less expensive than glutaraldehydes. such as laparoscopes. if not rinsed off. Glutaraldehydes. they can be chemically sterilized. which cannot be heated. Some high-level disinfectants will kill endospores after prolonged (10–24 hour) exposure. this residue can interfere (cause sticking) with the sliding parts of the laparoscope and cloud the lens. If objects need to be sterilized. Death of microorganisms occurs with dry heat by a process of slow destruction of protein. but dry-heat sterilization can be achieved with a simple oven as long as a thermometer is used to verify the temperature inside the oven. heat is absorbed by the exterior surface of an item and then passed to the next layer. Initially. Dryheat sterilization takes longer than steam sterilization. Sterilization takes place by soaking for at least 10 hours in 2–4% glutaraldehyde solution or at least 24 hours in 8% formaldehyde. CHEMICAL STERILIZATION An alternative to high-pressure steam or dry-heat sterilization is chemical sterilization (often called “cold sterilization”). A convection oven with an insulated stainless steel chamber and perforated shelving to allow the circulation of hot air is recommended.

a practice long since shown to be ineffective and unnecessary (Schmidt 1899). lowtemperature steam formaldehyde sterilizers that are . Commonly used liquid alcohol concentration of 70-80% due to higher concentrations less effective or lower. chlorine gas with water to produce ions that will the coagulation hipokloride proteins so cell membrane damage and enzyme inactivation occurred. When using either glutaraldehydes or formaldehyde. power works is the protein coagulation. ethyl alcohol is very effective at levels of 70% rather than 100% and this does not kill the spores. power works is to react with tyrosine. 4) Formaldehyde concentration of 8% is powerful enough to kill most microorganisms. There are. But alcohol can get rid of oil. Sterilization with chemicals used alcohol 70%. According to Gupte (1990). automatic. Alcohol 70% sprayed on praktikan hands and tools such as makropipet and micro-pipettes. 2) Chlorine. According to Volk and Wheeler (1988). the material becomes damaged when sterilized at high temperatures can be chemically sterilized using gas. One of the first uses of formaldehyde gas was to fumigate rooms. an amino acid or protein in emzim microorganisms. 70% alcohol can cause protein denaturation and koagulaasi.carcinogen (Rutala 1996). GAS STERILIZATION The use of formaldehyde gas for killing microorganisms was practiced before the turn of the century. 3) Iodine. 5) ethylene oxide gas. Sterilization with alcohol made on the process of making stock culture and isolation techniques. wear eyewear to protect from splashes. the gas is used mainly to sterilize materials made of plastic. According to Lay and Hastowo (1992). wear gloves to avoid skin contact. alcohol use on the skin when the ignition is too short to cause a lot of the effects of germicides and alcohol soon evaporated because it is easy to evaporate. Chemicals are often used include: 1) Alcohol. According to Gupte (1990). dust particles.Power works is related to the acids in microbial protein. and bacteria. limit exposure time and use both chemicals only in wellventilated areas (Clark 1983). however.

inside the barrel of a needle or laparoscope). In addition. Sterilization using ETO.and moisture-sensitive surgical instruments. regular exposure to low levels (greater than 1 part per million) may produce harmful effects in humans. the gas is irritating to the eyes and mucous membranes. a toxic product. UV light does not penetrate dust. patients and the environment. UV irradiation can interrupt transmission of airborne infections in enclosed indoor environments where living conditions are poor and people are crowded together. it requires sophisticated equipment and skilled staff specially trained for its safe use. is classified as a potential carcinogen as well as a mutagen. ethylene oxide (ETO) gas is used for sterilization of heat. Although in theory intense UV light can be both bactericidal and viricidal. ETO is hazardous to healthcare workers. it cannot be used to sterilize water. The acid is rapidly effective against all microorganisms. This is because the UV rays can kill only those microorganisms that are struck directly by UV light beams. in practice only limited disinfection of instruments can be achieved. Because ETO is moderately toxic when inhaled.effective and can be used to process heat-sensitive instruments and plastic items. eyes and respiratory tract. however.g. such as plastic devices and delicate instruments.. disposing of it is difficult (Gruendemann and Mangum 2001). ULTRAVIOLET LIGHT STERILIZATION Ultraviolet (UV) light has been used to help disinfect the air for more than 50 years (Morris 1972). any microorganisms present will not be killed (Gruendemann and Mangum 2001). Finally. because formaldehyde vapors are irritating to the skin. As mentioned previously. OTHER CHEMICAL STERILANTS  Paracetic acid (peroxyacetic acid). because ethylene oxide.and residual ETO on instruments can cause skin injuries and inflammatoryreactions in patients. For surfaces that cannot be reached by the UV rays (e. despite manufacturers= claims. For example. the use of formaldehyde in this form should be limited. mucous or water. Because UV irradiation has very limited energy. Therefore. is a more complicated (requires a 2-hour exposure time and a long aeration period) and expensive process than either steam or dry-heat sterilization. Moreover. organic matter does not diminish its activity and it . making it impractical for use in many countries (Gruendemann and Mangum 2001). In the United States and several other countries.

This solid polymer of formaldehyde may be vaporized by dry heat in an enclosed area to sterilize objects. Additionally the tool is also used to sterilize media.) Before the start. time and the lab. .Noteworthy during these devices operate is: Who writes the user (name.  Sterilization of tools a. mortar pestle and others. It is usually used for sterilizing different types of endoscopes and other heat-sensitive instruments. This method can sterilize items in less than 1 hour and has no harmful by products. It can also be used to sterilize soil or compost to be used for medium plants. Sterilization stage is actually quite short with a temperature of 121 degrees Celsius for 15 minutes.decomposes into safe products. It does not penetrate well. and cannot be used on paper or linen. A specialized sterilizer is required for performing gas plasma sterilization. Autoclave Autoclave used to sterilize equipment such as biotech tip. the overall time from initial heating (temperature rise) until cooling (temperature drop) could reach approximately 2 hour's. may be well suited for sterilizing endoscopes and other heatsensitive instruments. When diluted. The picture above is an example of the simplest type of autoclave. It aims to further refine the process of sterilization. called “selfsterilization”. The working principle tool is the same as the working principle of the steamer (a simple tool to cook rice) only has the pressure to produce higher heat. however.  Paraformaldehyde. both jelly or liquid media. e-tube. it is very unstable and must be used with a specially designed automatic sterilizer (APIC 2002). This technique.  Gas plasma sterilization (hydrogen peroxide based). However.

always wear heat resistant gloves. if the laboratory is located at a certain height. To the workings of the use of autoclaves have been made before. The principle of the workings of the autoclave As was explained in part on the introduction chapter tools. then the pressure was raised to 20 psi so that the temperature reached 1210C to boil water.8 0F because water boils at that temperature if it is used a pressure of 15 psi. Autoclave should not be opened before the pressure reaches 0 psi. then the pressure setting needs to be set again. autoclave is a device for memsterilkan wide range of tools & materials that use a pressure of 15 psi (1. All forms of life will die if it is boiled at a temperature of 1210C and a pressure of 15 psi for 15 minutes. Remember this event is only applicable to sea-level.02 atm) and temperature of 1210C. the heat source is turned off and allowed to drop slowly until the pressure reaches 0 psi. Reasons to use temperature is 1210C or 249. Then the sterilization process begins and the timer starts counting timer. To pressure 0 psi at sea level altitude (sea level) water boils at a temperature of 1000C. do not open the autoclave prior to cold temperatures (below 60 degrees centigrade ). the water in the autoclave over time will boil and steam is formed that fills the air pressing autoclave. whereas for the autoclave is placed on the same height. At the time reached the appropriate pressure and temperature. using a pressure of 15 psi then the water will memdididh at 121 0C temperature. Temperature and high pressure are given to media sterilized tools and gives greater strength to kill the cell compared with hot air. Usually used for media to sterilization 1210C temperature and pressure of 15 lb/in2 (SI = 103. After the sterilization process is complete. For example autoclave is placed at an altitude of 2700 feet above sea level. At the time of the heat source is turned on. fill the water according to the size specified before the start. After all the air in the autoclave was replaced with water vapor. ..4 Kpa) for 15 minutes. valve steam / air is closed so that air pressure in the autoclave increased.

Material can not stand the heat such as serum. Sterilization procedures with radiation (with Laminar Air Flow) . the remaining empty space dibirkan. Hands spray. antibiotics.5 should not be sterilized by autoclaving . browning (medium to brown) and the destruction of the substrate can be performed following precautions: . Procedures 1. Tools and Materials Tools: Autoclave. If sterilize media 1L 2L erlenmeyer accommodated on the sterilization time is set to 30 minutes. Materials: Alcohol. After the sterilization process and then grown on media. such as SDS To prevent the occurrence of precipitation.Paelarut organic. vitamins. Sterilization procedures work area 2. Oven. normally these microbes are commercially available in the form of spore strips. D. and enzymes .Compounds separated with sterilized mineral salts in order .Media that has a pH> 7. Ose pin. alumunium foil. cotton.Buffer ith the content of detergents. Sterilization by dry heat procedure (with oven) 5. Cawan Petri. E. Some media or materials that are not sterilized by autoclaving is: . Spore strips of paper are included in the autoclave and sterilized.Do not sterilize the solution to a pH <6.Separate sterilized phosphate compounds with amino acids (Peptone) or other mineral salts. . Erlenmeyer. Sterilization Procedures by combustion (flame Bunsen) 3. If the media remains clear then shows the autoclave has been working well.Glucose sterilized separately with amino acids (Peptone) or phosphate compounds . Bunsen.0 Erlenmeyer maximum media should only be filled ¾ of the total volume. such as phenol . Sterilization by moist heat procedure (by autoclave) 4. etc.To detect that the autoclave working perfectly usable pengguji microbes that are thermophilic and have endospores of Bacillus stearothermophillus.

2) Set the equipment to be sterilized in an autoclave baskets in a position where approximately the entire surface of the tool can be reached by steam in an autoclave.F. 4) Spray also our hands to be sterile. then the exhaust hole is closed. 3) Close the autoclave with a tight. Moist Heat Sterilization Procedures (With autoclave) 1) Ensuring enough water in the autoclave (high water 2 cm) below the bottom of the basket) or as much as 3-5 liters. 2) Sterilize a needle with the burning tip of the needle loop made of metal in the Bunsenflame.Or touched the tip of loop on the surface of the media until you hear a "ches". 5) Putting the tools to be used on a desk. . do not immediately open the lid of the autoclave. 6) Spray the back of our hands when they wanted to use those tools. c. Sterilization Procedures Work Area 1) Insert the alcohol solution with levels of 70% into a spray bottle. 20 minutes and already sounded the alarm sign is completed. 2) Spray the air around the work area with alcohol. fuel andspark glowing needle until it propagates to the base of the metal needle. Holding the needle with a slightly upright position on the fire ( 45 ). 3) Spray the desk with alcohol and smooth with a paper tissue. 3) When you're done. b.5) After the autoclave working for But wait until the pointer points to the 0 pressure. which is characterized by the release of steam from the exhaust hose is accompanied by a hissing sound. wait a few seconds until the temperature returns to normal OSE. Sterilization Procedures With Arson (Bunsen flame) 1) Turn on the Bunsen burner by opening the lid. then set the controller time at number 20 (20 minutes) and make sure the exhaust controller in an open and drain the controller in a closed state. 4) After the steam rises. Observation Result a.

The Radiation Sterilization Procedures (With Laminar Air Flow) 1) Wiping surface with alcohol 70% laminar. khususnya percobaan dan pengamatan mikroorganisme. 3) Close the curtains as close as possible until felt there would be no light coming out. 2) Close the oven and set the temperature controller on the numbers 160-1800 C for 1-2 hours. 4) Turn on laminar (the UV) at least 15 minutes. Untuk mengetahui teknik sterilisasi bahan-bahan yang digunakan dalam percobaan biologi. Sterilisasi Mekanik Sterilisasi mekanik. 12 Oktober 2011 di laboratorium microbiologi. 1. e. Sterilisasi yaitu suatu proses atau kegiatan membebaskan bahan atau benda dari semua bentuk kehidupan. Sterilisasi juga dapat diartikan sebagai proses untuk membunuh semua jasad renik yang ada. Filtrasi disini menggunakan suatu saringan yang berpori kecil sehingga mikroba dapat tertahan . Tujun dari pengamatan kali ini adalah untuk mengetahui teknik sterilisasi alat-alat yang digunakan dalam percobaan biologi. Sterilisasi mekanik dilakukan secara filtrasi. khususnya percobaan dan pengamatan mikroorganisme. Dry Heat Sterilization Procedures (In Oven) 1) Unscrew the oven and insert the equipment from a glass that has been wrapped into the oven. Discussion Pengamatan teknik sterilisasi dilakukan pada hari Rabu. biasa dilakukan dengan filtrasi. 6) Open drapes and turn on the blower when it will work. fisik dan kimiawi. dan untuk sterilisasi kimiawi dapat dilakukan menggunakan senyawa disinfektan. 5) Turn off the UV button when finished. sehingga jika ditumbuhkan di dalam suatu medium tidak ada lagi jasad renik yang dapat berkembang. G. 7) If less light. 2) Putting the tools and materials to be sterilized into the laminar. sterilisasi fisik dilakukan dengan pemanasan dan penyinaran. Pada prinsipnya sterilisasi dapat dilakukan dengan 3 cara yaitu secara mekanik. turn on the light.d.

poliester. yaitu: a. dll. Salah satu alat yang digunakan dalam pembakaran langsung adalah Bunsen. Sterilisasi panas kering cocok untuk alat yang terbuat dari kaca misalnya erlenmeyer. selulosa nitrat. Sterilisasi Fisik Pada sterilisasi fisik dapat dilakukan dengan 2 sistem. c. Pemanasan basah biasanya dilakukan didalam autoklaf atau aterilisator uap yang mudah diangkat dengan menggunakan uap air jenuh . Bila sudah selesai. Pemijaran (dengan api langsung): membakar alat pada api secara langsung. Prosedur Sterilisasi Panas Kering dengan Oven adalah sebagai berikut: 1) Membuka tutup oven dan masukkan peralatan dari gelas yang sudah dibungkus ke dalam oven. yaitu system pemanasan dan penyinaran. polimer akrilik. 2) Mensterilkan jarum ose dengan membakar ujung jarum yang terbuat dari logam pada api bunsen. pemanasan basah : menggunakan autoklaf. d. Atau menyentuhkan ujung ose pada permukaan media hingga terdengar bunyi ”ches”. 2. politef. dan berbagai tipe bahan lain termasuk memban logam. pinset. bakar jarum hingga berpijar dan pijaran tersebut merambat 3) hingga pangkal bagian logam jarum. Bahan yang mengandung air lebih tepat menggungakan metode ini supaya tidak terjadi dehidrasi. System pemanasan sendiri terdiri dari 4 macam. polivinil klorida. menunggu beberapa detik hingga suhu ose kembali normal. nilon. polikarbonat. Ukuran nominal pori penyaring 0.pada saringan tersebut. Memegang jarum dengan posisi agak tegak di atas api ( 45). contoh alat : jarum inokulum. vinil.2 μm atau kurang dan penyaring dibuat dari berbagai jenis bahan seperti selulosa asetat. tabung reaksi dll. Panas kering: sterilisasi dengan oven kira-kira 60-180 0C. adapun prosedur kerjanya dalam mensterilkan jarum ose adalah sebagai berikut: 1) Menyalakan pembakar bunsen dengan cara membuka tutupnya. Pemanasan basah adalah sterilisasi panas yang digunakan bersama-sama dengan uap air. 2) Menutup oven dan mengatur pengontrol suhu pada angka 160-180C selama 1-2 jam. batang L. b. florokarbonat. Uap air panas: konsep ini mirip dengan mengukus.

Sebelum melakukan sterilisasi cek dahulu banyaknya air dalam autoklaf. Masukkan peralatan dan bahan. serta yang digunakan untuk destruksi atau sterilisasi kotor.bertekanan pada suhu 1210C selama 15 menit (Hadioetomo. Nyalakan autoklaf. 4. Kemudian klep . Tutup autoklaf dengan rapat lalu kencangkan baut pengaman agar tidak ada uap yang keluar dari bibir autoklaf.02 atm) dan suhu 1210C. Jika mensterilisasi botol bertutup ulir. Autoklaf yang ada di laboratorium terbagi menjadi dua yaitu yang digunakan untuk sterilisasi alat dan medium(media) yang akan dipakai. Autoklaf adalah alat untuk mensterilkan berbagai macam alat & bahan yang menggunakan tekanan 15 psi (1. Cara pemanasan basah dapat membunuh jasad renik atau mikroorganisme terutama karena panas basah dapat menyebabkan denaturasi protein. diatur timer dengan waktu minimal 15 menit pada suhu 121oC. termasuk enzimenzim didalam sel (Fardiaz. Rongga di dalam autoklaf tidak boleh terlalu penuh diisi dengan benda-benda yang akan disterilisasikan agar dapat terjadi aliran uap yang cukup baik. 2. maka dapat ditambah air sampai batas tersebut. Klep pengaman jangan dikencang-kan terlebih dahulu. Cara Penggunaan autoklaf : 1. 5. 3. untuk menghindari terbentuknya kerak dan karat. maka tutup harus dikendorkan. 1992). Gunakan air hasil destilasi. Tunggu samapai air mendidih sehingga uapnya memenuhi kompartemen Autoklaf dan terdesak keluar dari klep pengaman.02 atm) dan suhu 1210C. Jika air kurang dari batas yang ditentukan. Autoklaf adalah alat untuk mensterilkan berbagai macam alat & bahan yang menggunakan tekanan 15 psi (1. 1985).

Kerusakan tersebut dapat diperbaiki bila disinari dengan berkas yang mempunyai gelombang yang lebih panjang. Jika bahan sudah mengeras khususnya pada medium padat. Setelah bahan atau medium sudah steril dengan autoclave. 6. Sinar Ultra Violet ini berfungsi untuk membunuh mikroba yang menempel pada permukaan interior Safety Cabinet. Daya kerjanya adalah absorbsi oleh asam nukleat tanpa menyebabkan kerusakan pada permukaan sel. tetapi ada hal lain yang masih harus dilakukan saat mengunakan bahan. seharusnya alat-alat tersebut dimasukan kedalam oven untuk dikeringkan.pengaman ditutup (dikencangkan) dan tunggu sampai selesai. Sterilisasi Kimia . Jika alarm tanda selesai berbunyi. Kemudian klep-klep pengaman dibuka dan keluarkan isi autoklaf dengan hati-hati. Sinar ultra violet dengan panjang gelombang yang pendek memiliki daya antimikrobial yang sangat kuat. Sedangkan untuk teknik penyinaran adalah menggunakan sinar UV. dadulu dalam hot plate sebelum Hal ini berfungsi untuk menghomogenkan suatu larutan dengan pengadukan. Penghitungan waktu 15’ dimulai sejak tekanan mencapai 2 atm. bahan dapat disimpan dalam lemari es khususnya untuk bahan yang mengandung unsure khusus agar tidak rusak. Oleh karena itu. 3. bahan bisa dipanaskan terlebih dipindahkan. Tapi hal ini tidak dilakukan karena waktu yang diperlukan tidak mencukupi. Pelat (plate) yang terdapat dalam alat ini dapat dipanaskan sehingga mampu mempercepat proses homogenisasi dan juga dapat digunakan untuk mencairkan media padat. Bahan yang sudah ada dalam autoclave sekarang sudah steril. Hal tersebut adalah menjaga tempat pembuatan atau pemindahan medium ke wadah yang lain missal dari elemeyer kedalam cawan Petri. maka tunggu tekanan dalam kompartemen turun hingga sama dengan tekanan udara di lingkungan (jarum pada preisure gauge menunjuk ke angka nol). Karena sterilisasi ini menggunakan uap air panas sebagai agen pensterilnya sehingga saat alat-alat dikeluarkan dari autoklaf banyak sekali tetesan uap air yang mengenai alat.

Laminary Air Flow adalah alat yang berfungsi untuk meniupkan udara steril secara kontinue melewati tempat kerja sehingga tempat kerja bebas dari debu dan spora-spora yang mungkin jatuh ke dalam media. Namun alkohol dapat menyingkirkan minyak. adapun proses penggunaannya adalah sebagai berikut: 1) Memasukkan larutan alkohol dengan kadar 70% ke dalam botol semprot. Salah satu kegunaan dari alcohol ini adalah untuk sterlisasi area tempat kerja. Menyemprot kembali tangan kita ketika hendak menggunakan alat-alat tersebut. etil alkohol sangan efektif pada kadar 70 % daripada 100 % dan ini tidak membunuh spora. Menurut Gupte (1990). disemprot terlebih dahulu dengan alcohol 70% atau spiritus. waktu pelaksanaan pemindahan bahan maupun proses penanaman.Sterilisasi secara kimiawi biasanya menggunakan senyawa disinfektan. kita juga harus mempertimbangkan faktor lingkungan yang ada. Menyemprot meja kerja dengan alkohol dan ratakan dengan kertas tissue. Saat mengunakan Laminary Air Flow. alkohol bila digunakan pada kulit kontaknya terlalu pendek untuk menimbulkan banyak efek germisida dan alkohol segera menguap karena sifatnya mudah menguap. Jika lingkungan tidak steril. Sterilisasi dengan bahan kimia digunakan alkohol 70 %. Alat-alat yang dimasukkan ke dlam laminar air flow cabinet. partikel debu. Senyawa disinfektan yang biasa digunakan yaitu alcohol. Menyemprot udara di sekitar area kerja dengan alkohol tersebut. Alkohol 70 % disemprotkan pada tangan praktikan dan alat-alat seperti makropipet dan mikropipet. Menurut Volk dan Wheeler (1988). Pada saat pemindahan bahan. Sterilisasi dengan alkohol dilakukan pada proses pembuatan kultur stok dan teknik isolasi. Kemudian menyiapkan semua alat-alat steril yang akan dipergunakan. maka bahan dapat terkontaminasi oleh bakteri atau mikroorganisme yang ada di udara atau dilingkukan itu sendiri. Menyemprot juga tangan kita agar steril. dan bakteri. 2) 3) 4) 5) 6) alkohol 70 % dapat menyebabkan denaturasi protein dan koagulaasi. Meja dan dinding dalam Laminary . hal yang harus dilakukan sebelum mengunakannya adalah menyalakan lampu UV. Meletakkan alat-alat yang akan digunakan pada meja kerja. sebelum laminar air flow digunakan dan hindarkan sinarnya dari badandan mata agar sel tidak mengalami mutasi. Menurut Gupte (1990). Untuk itulah kita harus mengunakan Laminary Air Flow. minimum selama 30 menit.

.Using the compound disinfectant (alcohol) .Gramedia. in Infection Prevention in Surgical Settings.Irradiation I. Departemen Pendidikan dan Gruendemann BJ and SS Mangum.Autoclave . dan pemindahan bahan. 3. Itu semua adalah teknik pensterilan bahan mulai dari pembuatan. 1985. Chemistry Sterilization.Air Flow juga harus disemprot dengan alkohol 70% atau dengan spiritus terlebih dahulu untuk mensterilkan Laminary Air Flow. such as: . Materials sterilization techniques. Srikandi. Mikrobiologi Dasar dalam Praktek. Conclution 1. pp 32–35. 1992. 2001. Tools sterilization techniques.Jakarta. 2. maka bahan dapat dimasukan kedalam Laminary Air Flow untuk proses pemindahan atau penanaman. WB Saunders Company: Philadelphia. Physical. that is mechanic.Direct Heating . Reference Fardiaz. PT. penyimpanan. Sterilization is intended to prevent contamination of equipment and materials used in microbiological testing by bacteria and viruses from the surrounding environment.Dry Heat (with oven) .Filtration . R. H. Ultraviolet irradiation and lights. Setelah semuanya steril. Sterilization of tools and materials can be done with 3 ways. Sterilization in general is a method of destruction of all forms of living organisms.Mikrobiologi Pangan.Hot wet (using an autoclave) . Hadioetomo.S. such as: .Steam hot water 4.

M. Institut Pertanian Bogor. Surg Gyne Obstet 69: 738–744. Studies on chemical sterilization of surgical instruments. 1988. Clinical Infectious Diseases 32(9): 1348– 1356. Mikrobiologi Dasar. Rutala WA and DJ Weber.A. W. Spaulding EH 1939. Volk. dan Wheeler.Kebudayaan. Creutzfeldt-Jakob disease: recommendations for disinfection and sterilization. Penerbit Erlangga. PAU Pangan dan Gizi.F. 2001. .