1. Wet Steam Sterilization
2. Sterilization dry Warming
Using an autoclave, in principle, sterilization autoclave uses heat and pressure of water vapor. Temperature sterilasi typically 121 C, the pressure normally used between 15 to 17.5 psi (pounds per square inch) or 1 atm. The duration of sterilization depends on the volume and type. The tools and sterilized water for 1 hour, but the media between 20-40 minutes depending on the volume of material sterilized. Sterilization of media for too long causes:
1. The decomposition of sugar
1. The decomposition of sugar
2. Degradation of vitamins and amino acids.
3. Inactivation cytokinin zeatin riboside.
4. PH changes for berakibatkan depolymerization
3. Inactivation cytokinin zeatin riboside.
4. PH changes for berakibatkan depolymerization
Autoclave portable gas or electricity in general use steam from the heating source of water is added into the autoclave, while the large autoclave at commercial laboratories typically use steam from a central boiler.
Parts of the autoclave:
1. Pan out.
2. Pan in place put the bottle with grooves where the steam channel.
3. Close the bookmark along with pressure and steam lines.
4. Vaporization valve.
5. Lockout or clamps
Parts of the autoclave:
1. Pan out.
2. Pan in place put the bottle with grooves where the steam channel.
3. Close the bookmark along with pressure and steam lines.
4. Vaporization valve.
5. Lockout or clamps
The weakness of this autoclave is that the necessary safeguards and heat settings manually, during the sterilization done. But this autoclave has advantages: simple, relatively cheap price, irrespective of the flow of electricity is often a problem for countries that are developing, as well as faster than the electric autoclave size and similar.
Autoclave which is more comprehensive use of energy sources of electricity. Appliance is equipped with a timer and thermostat. If the automatic regulator is going well. Then the autoclave can be run while doing other work. The downside is that if one regulator is not working, then the preparatory work the media to be in vain and possibly cause total failure of the autoclave. As a source of steam, also derived from water that is added into the autoclave and bring to a boil
Autoclave which is more comprehensive use of energy sources of electricity. Appliance is equipped with a timer and thermostat. If the automatic regulator is going well. Then the autoclave can be run while doing other work. The downside is that if one regulator is not working, then the preparatory work the media to be in vain and possibly cause total failure of the autoclave. As a source of steam, also derived from water that is added into the autoclave and bring to a boil
Portable Media use autoclave sterilization
(Warm-use of fire)
1. Fill outer pan with water, if possible with distilled water to avoid precipitation of Ca commonly found in tap water, 1 liter to a small autoclave, and 1.5 liters for large autoclave.
2. Bottles of media to be sterilized, is inserted into the panel-in. Arrange the bottles until it reaches the surface of the panel.
3. Set the pan position with respect to the flow channel where steam is present in the surface of the pot lid and the outer circle
(Warm-use of fire)
1. Fill outer pan with water, if possible with distilled water to avoid precipitation of Ca commonly found in tap water, 1 liter to a small autoclave, and 1.5 liters for large autoclave.
2. Bottles of media to be sterilized, is inserted into the panel-in. Arrange the bottles until it reaches the surface of the panel.
3. Set the pan position with respect to the flow channel where steam is present in the surface of the pot lid and the outer circle
4. Cover tightly. (Tighten the lock without using tools)
5. Let the steam valve is open expenditure.
6. Place the autoclave on the gas stove or a Bunsen burner.
7. Heat to boiling water in an autoclave and steam coming out of valve steam expenditure.
8. Let the steam out for 5 minutes (minimum), to remove the air out of air trapped in the autoclave.
9. Close the steam valve expenditure.
10. Observe the increase of temperature and pressure.
11. Once the pressure reaches 15 psi, fire stove understated.
12. Keep the pressure of 15 psi this situation by adjusting the size of the burner manually. During sterilization, do not leave the autoclave and the other room doing other things, because the pressure can be increased up to cross the line. This situation is dangerous and can cause damage to equipment.
13. After the sterilization time is reached, turn off the stove fire.
14. Steam released a little bit by adjusting the steam valve expenditure (open a little).Never open the valve and let the steam out at once. This causes the media or the water bubble up
15. After the pressure drops to 0, open the lock and remove the pan containing the media
5. Let the steam valve is open expenditure.
6. Place the autoclave on the gas stove or a Bunsen burner.
7. Heat to boiling water in an autoclave and steam coming out of valve steam expenditure.
8. Let the steam out for 5 minutes (minimum), to remove the air out of air trapped in the autoclave.
9. Close the steam valve expenditure.
10. Observe the increase of temperature and pressure.
11. Once the pressure reaches 15 psi, fire stove understated.
12. Keep the pressure of 15 psi this situation by adjusting the size of the burner manually. During sterilization, do not leave the autoclave and the other room doing other things, because the pressure can be increased up to cross the line. This situation is dangerous and can cause damage to equipment.
13. After the sterilization time is reached, turn off the stove fire.
14. Steam released a little bit by adjusting the steam valve expenditure (open a little).Never open the valve and let the steam out at once. This causes the media or the water bubble up
15. After the pressure drops to 0, open the lock and remove the pan containing the media
For culture media that do not contain ingredients that Heat-labile, sterilization is done by autoclave at a temperature of 121oC, pressure between 15 psi or 1 atm with a time between 20-25 minutes depending on the volume of container and the volume of media.To 15-50 ml of medium in test tubes or small sized bottles 50-100 ml, sterilization performed at a pressure of 15 psi with a time of 20 minutes. For the 20 bottles of 1 liter volume requires a longer time to 34 minutes, 10 bottles of 2 liters volume takes 37 minutes, 5 bottles of 4 liters of time spent on 52 minutes. With a longer time. In distilled water and sterilizing the media, after sterilization the desired time is reached, the autoclave pressure should not be reduced abruptly. When the pressure is suddenly reduced, the liquid inside boils and bubbles (bubbled up).
For materials that heat-labile in the form of solutions, sterilization performed by filtering the solution through a filter that has a pore size um 0:20 to 0:22. The diameter of the filters vary depending on the volume of solution to be sterilized. For the solution volume 10 ml, used filters mounted on the tip of a syringe. Materials are heat labile, among others: GA3, Thiamin-HCL, Ca-panthothenate, Antibiotics: carbenocilin.
For materials that heat-labile in the form of solutions, sterilization performed by filtering the solution through a filter that has a pore size um 0:20 to 0:22. The diameter of the filters vary depending on the volume of solution to be sterilized. For the solution volume 10 ml, used filters mounted on the tip of a syringe. Materials are heat labile, among others: GA3, Thiamin-HCL, Ca-panthothenate, Antibiotics: carbenocilin.
Sterilization Equipment Culture
1. Clean bottles were given a few drops of distilled water and cover with paper or aluminu
1. Clean bottles were given a few drops of distilled water and cover with paper or aluminu
aluminum foil (not too tight when using the al-foil). For the bottles that have a lid that autoclaveable, do not close too fast, because the expansion occurred during heating.
2. The tools need to be sterilized before planting are: tweezers, scissors, scalpel handles, filter paper, petri-dish, empty bottles, needles and pipettes.
3. The tools and filter paper carefully wrapped with thick paper or placed in a stainless steel tray and the tray is wrapped with thick cloth before it is inserted in an autoclave.Aluminum foil is not recommended as a wrap, as moisture can not enter into the bundle. Sektio tools such as tweezers, scissors, scalpel handles, and the needle, wrapped in paper or a paper coffee straw. Avoid the use of Al-foil as moisture is difficult to get into the package so that sterilization is less effective.
4. Petri-dish will be sterilized, also wrapped in paper or a paper coffee straw.
5. The temperature used for sterilization of the culture bottle is empty and the tools that will be used to plant explants, is 121o C at a pressure of 15 psi (pounds per square inch) or 1 atm for 30-60 minutes. Sterilization timing begins after the desired pressure and temperature is reached.
The tools used when planting, must be in sterile condition. Metal tools and glass can be sterilized in an autoclave. Cropping tool such as tweezers and scissors can also be sterilized by burning or by heating in bacticinerator. Especially for the scalpel, the handle can be sterilized by heating, but the blade (blade) can be blunt when heated in high temperatures. Therefore, for the way the blade is recommended sterilization by immersion in alcohol or chlorine solution.
2. The tools need to be sterilized before planting are: tweezers, scissors, scalpel handles, filter paper, petri-dish, empty bottles, needles and pipettes.
3. The tools and filter paper carefully wrapped with thick paper or placed in a stainless steel tray and the tray is wrapped with thick cloth before it is inserted in an autoclave.Aluminum foil is not recommended as a wrap, as moisture can not enter into the bundle. Sektio tools such as tweezers, scissors, scalpel handles, and the needle, wrapped in paper or a paper coffee straw. Avoid the use of Al-foil as moisture is difficult to get into the package so that sterilization is less effective.
4. Petri-dish will be sterilized, also wrapped in paper or a paper coffee straw.
5. The temperature used for sterilization of the culture bottle is empty and the tools that will be used to plant explants, is 121o C at a pressure of 15 psi (pounds per square inch) or 1 atm for 30-60 minutes. Sterilization timing begins after the desired pressure and temperature is reached.
The tools used when planting, must be in sterile condition. Metal tools and glass can be sterilized in an autoclave. Cropping tool such as tweezers and scissors can also be sterilized by burning or by heating in bacticinerator. Especially for the scalpel, the handle can be sterilized by heating, but the blade (blade) can be blunt when heated in high temperatures. Therefore, for the way the blade is recommended sterilization by immersion in alcohol or chlorine solution.
2. Autoklaf
Autoclave yaitu alat untuk mensterilkan berbagai macam alat dan bahan yang digunakan dalam mikrobiologi menggunakan uap air panas bertekanan. Tekanan yang digunakan pada umumnya 15 Psi atau sekitar 2 atm dan dengan suhu 121oC (250oF). Jadi tekanan yang bekerja ke seluruh permukaan benda adalah 15 pon tiap inchi2 (15 Psi = 15pounds per square inch). Medium yang akan disterilkan ditempatkan di dalam autoclave selama 15-20 menit, hal ini bergantung pada banyak sedikitnya barang yang perlu disterilkan. Medium yang akan disterilkan ditempatkan dalam beberapa botol yang agak kecil daripada dikumpul dalam satu botol yang besar. Setelah pintu autoclave ditutup rapat, barulah kran pada pipa uap dibuka dan temperatur akan terus-menerus naik sampai 121oC (Dwidjoseputro, 1990)
 |
| Add caption |
1. Tombol pengatur waktu mundur (timer)
2. Katup pengeluaran uap
3. pengukur tekanan
4. kelep pengaman
5. Tombol on-off
6. Termometer
7. Lempeng sumber panas
8. Aquades (dH2O)
9. Sekrup pengaman
10. batas penambahan air
Cara menggunakan autoclave:
a. Sebelum melakukan sterilisasi cek dahulu banyaknya air dalam autoklaf. Jika air kurang dari batas yang ditentukan, maka dapat ditambah air sampai batas tersebut.Gunakan air hasil destilasi, untuk menghindari terbentuknya kerak dan karat.
b. Masukkan peralatan dan bahan. Jika mensterilisasi botol beretutup ulir, maka tutup harus dikendorkan.
c. Tutup autoklaf dengan rapat lalu kencangkan baut pengaman agar tidak ada uap yang keluar dari bibir autoklaf. Klep pengaman jangan dikencangkan terlebih dahulu.
d. Nyalakan autoklaf, diatur timer dengan waktu minimal 15 menit pada suhu 121oC.
e. Tunggu samapai air mendidih sehingga uapnya memenuhi kompartemen autoklaf dan terdesak keluar dari klep pengaman. Kemudian klep pengaman ditutup (dikencangkan) dan tunggu sampai selesai. Penghitungan waktu 15’ dimulai sejak tekanan mencapai 2 atm.
f. Jika alarm tanda selesai berbunyi, maka tunggu tekanan dalam kompartemen turun hingga sama dengan tekanan udara di lingkungan (jarum pada preisure gauge menunjuk ke angka nol). Kemudian klep-klep pengaman dibuka dan keluarkan isi autoklaf dengan hati-hati
Autoclave is a means to sterilize a variety of tools and materials used in microbiology using pressurized hot water vapor. The pressure used was 15 psi, or about 2 atm and a temperature of 121oC (250oF). So the pressure that work to the entire surface of the object is 15 pounds per inchi2 (15 Psi = 15pounds per square inch). Medium to be sterilized are placed in the autoclave for 15-20 minutes, this depends on the extent of goods that need to be sterilized. Medium to be sterilized are placed in several small bottles rather than collected in a bottle that big. After the autoclave door is closed tightly, then opened the steam valve on the pipe and the temperature will continue to rise until 121oC
Diagram of vertical autoclave
1. Key regulator timer (timer)
2. Valve vaporescence
3. pressure gauge
4. safety valve
5. The on-off
6. Thermometer
7. Plate heat sources
8. Aquades (dH2O)
9. Security Screws
10. limit the addition of water
How to use an autoclave:
a. Before performing the first check number of water sterilization in an autoclave. If water is less than the specified limit, then the water can be added to the distilled water tersebut. Used limit, to avoid the formation of crust and rust.
b. Enter the equipment and materials. When sterilizing bottles beretutup threaded, then the lid should be relaxed.
c. Close the autoclave with a safety meeting and tighten the bolt so that no steam coming out of the lips autoclave. Safety valves do not be tightened first.
1. Key regulator timer (timer)
2. Valve vaporescence
3. pressure gauge
4. safety valve
5. The on-off
6. Thermometer
7. Plate heat sources
8. Aquades (dH2O)
9. Security Screws
10. limit the addition of water
How to use an autoclave:
a. Before performing the first check number of water sterilization in an autoclave. If water is less than the specified limit, then the water can be added to the distilled water tersebut. Used limit, to avoid the formation of crust and rust.
b. Enter the equipment and materials. When sterilizing bottles beretutup threaded, then the lid should be relaxed.
c. Close the autoclave with a safety meeting and tighten the bolt so that no steam coming out of the lips autoclave. Safety valves do not be tightened first.
d. Turn on the autoclave, set the timer with a minimum of 15 minutes at a temperature of 121oC.
e. Wait samapai boiling water so the steam meets the autoclave compartment and pushed out of a safety valve. Then the safety valve is closed (tightened) and wait until the end. Calculation of time 15 'starting from the pressure reached 2 atm.
f. If alarm signal sounds finished, then wait until the pressure in the compartment down the same air pressure in the environment (in preisure gauge needle points to zero).Then the safety valves open and remove the contents of the autoclave with caution
e. Wait samapai boiling water so the steam meets the autoclave compartment and pushed out of a safety valve. Then the safety valve is closed (tightened) and wait until the end. Calculation of time 15 'starting from the pressure reached 2 atm.
f. If alarm signal sounds finished, then wait until the pressure in the compartment down the same air pressure in the environment (in preisure gauge needle points to zero).Then the safety valves open and remove the contents of the autoclave with caution
Tidak ada komentar:
Posting Komentar