‫محاضرة ‪3‬مايكرو عملي‬

Autoclave

✦ Use / used for sterilizing culture media, glassware, and other materials that
are not spoiled by moist heat .
✦principle /Moist heat & pressure.
 Incubator

 Use / used for providing microorganism with optimum temperature for living .
 Principle / electrical heat .
 Hot air oven

 Use / used for sterilizing glassware & other materials thar are spoiled by moist heat .
 Principle / Dry heat
 Inoculating loop

 Use/used for transferring and streaking cultures .

 Sterilized by burner .
 Vortex mixer

 Use/used for mixing liquids kept in test tube .

 Principle / electrically powered and moves the liquid in gyratory motion .
 Waterbath

 Use/used for heating and melting media, solutions and samples .

 Principle /Water tepemrature below 100C .
 Heating mantle

 Use / used to heat or melt sample and reagents , also used to water heating in
beaker .
 Principle / indirect heat
 Hot plate

 Use/ used to heating and agitation .

 Principle /magnetic beads.
 Uv Chamber

 Use/ used for analyzing fluorescent materials

 Principle /long and short wavelength Uv radiation .
 Inoculation chamber

 Use / used for culture transferring and plating

 Principle /UV light
 PHmeter

 Use / used for measuring hydrogen ion concnteratio.

 Colony counter
Use / used foer countering microbial colony .
Principle / backlight source, gridlines, magnifying lense and sensor .
 Microscop

 Use /used for observing microscopic items such as cells, crystals and cell organelles .
 Refrigerator

 Use /used for storing culture, media, many sensitive materials .

 Principle /elecrically powerd anduse ammonia as refrigerant .
 Bunsen burner

 Use / used for sterilizing loop, plating out culture and transferring culture .
 Spirit lamp

 Use/The same as burner but lamb is portable abs used as fuel) produces free smock
flam .)
 Lab balance

 Use / used to measure small mass in sub_milligram unit .

 Distillator

 Use / used to purfication and condensation of substance .

 Centrifuge

 Use / used for seperating liquides and samples of different densities .

🙂 ‫هذا مابي صوره بس اكيد شايفينه‬
 )‫(التعقيم‬4 ‫محاضرة‬

Sterelization methods
✦ Physical method

✦mechanical method
✦chemical method

Q/Differance between sterilization and disinfection ?

 Physical agents
✦Heat
✦Radiation

Q/Factors that influncing on heat sterilization ?

•Types of heat dry or moist .
•Temperature and time .
•Species of microorganism s
•No .Of microorganism presented
•Type of material to be sterilized .
‫ذني مهمات هواي سألت بيهن‬
 Types of dry heat

•Flaming / used for sterilizingl oop,needls, wires, glassslids, coverslips .

•Incineration/used for destroying contaminated materials .
•Hot air oven) 160C for 2hours)
‫واالستخدام ومبدأ العمل نعرفه‬
Q/Disadvantages of oven?
•Long sterilization cycle
•Require accurate pyrometer
•Damage to heat sensitive materials
 Moist heat

•Pasturazation / Slow method /63C for 30min

Fast method /72Cfor 15 sec followed by sudden cooling 13C or lower .
Ex. Pasteurization of milk .

•Boiling/ at 100C for 10_ 5min used to steril water .

Adding 2%NaHco3 to increasing afficacy .
•Tyndalization/ steaming at 100C for 30 min on each three days .
Used for cluture media containing sugar and gelatin .
•Autoclave/ 121C for 15 min at 15 psi
 Types of radiation

 Direct sun light / kill the vagitate organism rapidly .

 Non ionizing radiation (Uv light(used to sterilize air on surfaces) used in bacteriological
hood .)
 Ionizing radition/Gama rays, X_rays used to sterilize disposal such as gloves catheter .
 Infrared radiation/at 180C used to sterilize glassware .
 Mechanica method

 By filteration technique
 Used to sterilize sera, solutions of sugar , vaccines, toxins and antibiotics .

✦✦Disinfection / Ex. Alchohol

•Ethyl alchohol 70%
•Formaldehyde 10%
•phanols 5%
•hydrogen peroxide 6%_ 3
 )‫ (االوساط‬5 ‫محاضرة‬

 Culture media /Solutions containing all of nutriebts and physical parameters necessary for microbial growth .

Q/Classification of media
A/Based on composition
•Natural media consist of natural materials such as meat and peptone .
•Synthetic media consist of chemical materials
•Semisynthetic media consist synthetic materials in addition to natural materials .

Q//Difference between broth and groth

Broth/liquid media
Groth/solid media
 B_based on consistency

•Solid /contains 2% _ 1.5agar Ex .Blood agar

Used for determining colonies of bacteria .
•Semisolid/contains agar at 0.5or less concentration
Used for determining bacterial motility
•liquid/Contains spesific mount of nutrients without agar
Used for replication and adaptation of microorganism .
 C_Based on function

•Basic media /contains peptone, salt, water, source of carbon and source of nitrogen .
Ex .Nutrient broth, Nutrient agar
•Enrchied media /basic media that are supplmented with vitamin, body fluid, amino acid
and protein .
Ex .Blood agar and chocolate agar
 Types of hemolysis of bacteria

 Beta hemolysis is complete hemolysis ➡️clear zone around the colony

 Alpha hemolysis is partial hemolysis ➡️greenshied coloration of media
 Gamma hemolysis / No hemolysis
 Difference between differntial and
selective media

 Differential media / used for diffrentiatin between types of bacteria such as diffrentiation between lactose
_fermenting&non _lactose fermenting bacteria .
 ‫اذا اكو تخمر يكون الوسط لونه وردي واذا مااتخمر الالكتوز يكون الوسط‬
Colorless .

•Selectiv e media /used for growth of gram negative bacteria only such as macconky agar
T. B. C. S /selectiv e media , used for growth of v ibrio cholera
•S_S agar/differential ➡️differs between salmonella and shigella .

✦✦Differential media
•Macconky agar
•S_S agar
•Blood agar
 Mannitol salt agar

 Indicator media /contains ph indicator that change the colorr of media according to ph.)Mannitol salt agar)

 Special media / non differential and non selectiv e

 Such as Triple sugar iron agar /TSI/ contains lactose, glucose, sucrose
 Used to detect the ability of organisms to ferment glucose, sucrose, lactose and to produce H2S and gas .
 ‫اذا التيوب كله احمر ماكو تخمر‬
 ‫واذا من االعلى احمر وباالسفل اصفر معناها بس الكلكوز متخمر‬
 ‫واذا كله اصفر صاير تخمر للكلكوز واالكتوز والسكروز‬
 ‫واذا من االعلى اصفر وباالسفل احمر بس السكروز والالكتوزمتخمرات‬
 ‫اذا بيه فراغ يعني اكو غاز‬
 ‫واذا بيه سواد اكو‬
 H2s .
‫محاضرة ‪6‬‬
‫محاضرة ‪(7‬التصبيغ)‪.‬‬
Acid fast stain
Capsule stain
Amal Hussein
MARWA H.HASHEM