Sterilization
Preliminary
Sterilization is a process of freeing equipment or materials from
unwanted microorganisms. To achieve this goal, there are several types of
sterilization that can be selected and adjusted to the nature of the material to be
sterilized. General ways are done are:
a. Physical sterilization: sterilization by using heating, the use of UV rays, X-rays
and rays that are short wavelengths.
breakfast. Chemical sterilization: sterilization using chemicals, such as alcohol,
disinfectant, formalin solution, etc.
C.. Mechanical sterilization: sterilization using a filter or filter.
Sterilization with heating
a. Heating with dry air or wet air
This method is perfect for sterilizing heat-resistant tools or materials. In this
way, the temperature used is 170 oC and is done for 1 hour.
breakfast. Tindalisasi
This method is used to sterilize materials that cannot resist high temperatures.
In this way, the material and tool is heated for 30 minutes at 100 oC, and
repeated 3 times in a row, with a 1 day interval. Materials and tools are stored at
room temperature when not heated.
C.. Pasteurization
This method is used to sterilize food ingredients that will experience
denaturation at high temperatures. In this way, the tools and materials are
heated at a temperature of 60 - 80 oC for one hour 3 times in a row, with a 1 day
interval.
d. Sterilize directly.
This method is perfect for sterilizing the needle inoculation.
Hey. Sterilization with pressurized hot steam
This method is commonly used to sterilize high heat resistant tools and
materials and pressure. The tool used is Autoclave which can reach 121 OC
temperatures and 15 lbs pressure.
How to use Autoclave.
a. The contents of autoclave with 3-5 liters of water.
breakfast. Prepare a tool or material that will be sterilized on the Autoclave shelf.
C.. Enter the tools and materials on the shelf into the autoclave vessel, and cover
with a meeting (except air cleps).
d. Heat this autoclave and let all the air inside out (which is marked with the
release of drops of water through the air.
Hey. Close the air clap if all the air in Autoclave is out.
f. Heating continues until it reaches 121 oC and a pressure of 15 lbs, for 15
minutes.
g. Heating is stopped, the temperature is returned to room temperature and
pressure at point 0.
h. Never try to open Autoclave before
The pressure needle shows the number 0 (zero),
Because it's very dangerous !!!!!!!.
Note: When in Autoclave there:
a. 100% pure water vapor, the temperature achieved is 121 OC and a pressure of
15 lbs.
breakfast. 2/3 parts of water vapor and 1/3 of the air, then the temperature and
pressure achieved are 115 oC
and 15 lbs.
C.. 1/3 parts of water vapor and 2/3 air, then the temperature and pressure
achieved are 109 oC
and 15 lbs.
d. 100% air, then the temperature and pressure achieved are 100 oC and 15 lbs.
3.2. Sterilization with radiation
Done using short wave light, such as UV or CO 60 or CS 69 rays. In this lab a
solid medium sterilization will be carried out using UV light.
In doing autoclaving using this Autoclave do it in the following:
1. Fill in Chamber with water until the surface of water is slightly above the
surface
gold leaf.
2. Insert the material and tool into the kerajang in the Autoalve with rotate (Full)
Handle 1.
3. Turn on the machine with pressing power on.
4. Close the Autoclave meeting
5. Set temperature meters (4) to determine Autoclave milk (panel 4)
6. Adjust the old time of aucocklave (panel 5)
7. Close the steam panel 2 expenditure meeting)
8. Press Star (Panel 8)
Auclaving will run automatically, and the lights on the old panel time
Autoclaving will light up showing the autoclaving process (panel 7).
If the autoclaving process is complete, it will hear the sound
"Ti..ti ..... it ....".
After the process of autoclaving then do it as follows:
1. Press Stop (panel 9)
2. Open the steam spending tap (panel 2) until the steam all come out
3. Make sure the pressure is in the 'zero "machine (see Panel 3).
4. After the new zero terot autoclave machine can be opened and the material /
tool
Diligenced is issued and in a slash plager.
5. Turn off the Autoclave power engine.
A. Sterilization with UV
Tools
a. Petrol
breakfast. Medium to be upright.
C.. UV lights
d. Bunsen lights
Work
a. Disburse 3 medium tubes to upright in water bathing.
breakfast. Pour into 3 petri sterile dishes and let it freeze.
C.. Sinari was the first Petri dish with UV for 1 minute, a second cup for 3
minutes,
and the third cup is not illuminated (in an open state).
d. Incubation of Petri dishes for 24-48 hours at 37 oC, in reverse state.
Hey. Observe the growth of microbes in the three Petri dishes.
Sterilization with chemicals
Chemicals such as alcohol, formalin, carbol, licol, soap, and others can be used
to sterilize tools. In this lab the sterilization will be carried out using several
chemicals.
A. sterilization with alcohol
Tools
a. Petri sterile dish
breakfast. Medium na upright
C.. Pentul / small nail needle.
d. Alcohol with a concentration of 40%, 70% and 96%
Hey. Sterile water
f. Tweezers
Work
a. Melt the NA medium in water bathing and pour into the Petri dish. Let it
freeze on the tribe of the room.
breakfast. Soak the needle of pentul or nail in the alcohol solution above for 10
minutes.
C.. Wash the pentules with sterile water and place it on the medium surface in
Petri dishes.
d. Incubate at 28-30 oC for 24-48 hours.
Hey. Compare microbial growth around the Pentul needle. Which solution is
the most effective.
f. Do the same thing using 4% formalin, lisol and carbolic.
B. sterilization with soap
Tools
a. Petri sterile dish
breakfast. Medium na upright
C.. Soap with various trademarks.
Work
a. Melt 2 medium tubes upright in water bathing and pour into two petri sterile
dishes
breakfast. Appends a brother's finger that hasn't been washed on the medium
surface on the first Petri dish.
C.. Wash your fingers with certain brand soap as clean as possible and let it dry
by themselves (don't dry it with a lap). After drying, it is up to your finger on the
medium surface in the second Petri dish.
d. The incubation of the two petri dishes was 24-48 hours at 28-30 oC.
Hey. Compare microbial growth in the two petri dishes.
Sterilization with membrane filter
Materials that must not be heated, such as serum and some kind of sugar can be
stained using the filter membrane. In the field of microbiology, the most used
filter tool is a breederd filter, Chamberland filter, Seilz filter, etc. These filters
are made of special polymers with a diameter of 0.45 microns, so they can resist
the bacteria contained in the sample. In this practicum you will use a
membrane made of polysulfon, which is produced by Millipore.
Tools
a. Membrane polysulfon with a diameter of 0.22 microns (hydrophilic)
breakfast. A set of membrane trial tools complete with gas regulators.
C.. Petrol
d. McCartney bottle
Hey. Gun pipette complete with the tips
f. Medium na upright
g. Pollutam water
Work
a. Melt the medium na in water bath and leave it at a temperature of 40 oC.
breakfast. Filter this so that the water has been contaminated (water times)
using the polysulfon membrane.
C.. 1 ml of pipette. The water of this filter and put it in the petri dish
d. As a control, a 1 ml of water pipette that has not been filtered and placed in
the second Petri dish.
Hey. Pour each one tube medium na (temperature of 40 oC) into each petri dish
that has contained water samples.
f. Shake both cups in such a way that the sample becomes homogeneous with
the medium, and let it freeze.
g. The incubation of the two petri dishes at a temperature of 20-30 oC for 24-48
hours.
h. Observe the two dishes of Petri !! What conclusions can you take from this
experiment?
