Aseptic Technique - Microbiology
the high power objective magnifies
40 times so that is 400 with the eyepiece
Aseptic Technique is ?
A fundamental and important laboratory skill in the field of microbiology. Microbiologists use aseptic technique for a variety of procedures such as transferring cultures, inoculating media, isolation of pure cultures, and for performing microbiological tests.
Agar
A gelationous substance obtained from various kinds, used in media as a thicker in foods.
Safety on Culture
All culture media or trash that has come in contact with love bacteria or body fluids into the bio-hazard waste containers.
Bacterial Motility is
Chemotaxity and true motility is achieved by rotation of flagella.
Some inoculation techniques separate bacterial cells to form individual ?
Colonies
Nutrient Agar
Contains Nutrients that support growth of bacteria. Has a P.H. of 6.8 that is acidic
6 I's of Inoculation
Inoculation, Incubation, Isolation, Inspection, Information gathering, identifictation
Ubiquitous
It means that it is present , it appears or can be found everywhere.
During inoculation what instruments are used?
Loops and needles
Safety on Spills and Chemicals
Make sure to report all spills, even water. Keep containers away from edge of workbench. All Chemicals will stain clothing and tarnish jewelry.
Safety Question
Never taste a chemical. Only wafting of vapors towards your nose with your hands is allowed when told so by the instructor
Use ONLY fine adjustment to focus when using high power or oil.
Never use the coarse adjustment.
Why is aseptic technique necessary/important?
To keep infectious microorganism from sterile surfaces or tissues. Helps prevent contamination of test and or experiments in a lab.
Safety Question Bunsen and Burns
Turn off bunsen burner or hot plate when not using. If you get burned you run cold water on burn area for 15 minutes. The cold water reduces pain and blisters.
How do you return the light microscope to the cabinet?
Turn off the light and allow time to cool off. But the scanning objective back into position, coil up the cord properly and cover with the dust cover.
When you enter the lab?
You wash your hands with antibacterial soap. Clean your area with antiseptic spray, and before you leave make sure to wash your hands again with antibacterial soap.
colonies
a group of cells that are growing together
bacterial colonies
are elevated, colorful, sticky and shinny
Gram + bacteria
are stained purple with crystal violet
Gram - bacteria
are stained red with safranin
Electron Microscopes
are used in a high-tech lab where thin samples are scanned using electrons generating a computer image
Fungal colonies
are usually bigger, grow fast, puffy and dry
identify ways to practice aseptic techniques ?
boiling water, using Bunsen burner, autoclave machine, wash hands, sterilize area, don't place items down on a non-sterile surface.
Under the lower power, what adjustment knob do you use?
both, coarse and fine adjustment knobs to focus
how do you see the specimen?
by turning on the light source and center the specimen in the bean of the light.
When carrying the microscope you
carry it with one hand grasping the arm and the other hand underneath the base.
these bacteria are typically treated with penicillin
gram + bacteria
these bacteria are hard to treat even with penicillin
gram - bacteria
Condenser on the light microscope
has no visual on the microscope it's self. it concentrates light source into a cone of light that passes through diaphragm.
Inoculation
introduction of a pathogen or antigen into a living organism. Usually it's a vaccination, immunization, and/or by injection
Media plate
is a petri dish that has agar applied on it.
Growth Medium
is a solid, liquid or semi- solid designed to support the growth of microbes or cells. It contains nutrients that support microbial growth. It uses agar as a solidifying agent. It can be selective due to nutrients, P.H. and antibiotics.
Light microscope
is an instrument that is used to see organisms that are not seen by the naked eye.
Petri-Dish know hows and what to do.
label base using tape. With a sharpie, put your name, the date and what is inside. Always store your dish upside down. (that way the humidity will not enter-fear with your experiment) Seal your dish with parafilm. It is a stretchy tape like material used to seal your dish with.
The red scanning objective
magnifies 4 times plus the eyepiece so 40 times
Be sure to always center the specimen in your field aka the slide, of our view before rotating the
nose piece to change objectives.
The part that you look trough on the top of the microscope is called the
ocular eyepiece and it magnifies 10 times
Sub culturing (such as streak plating and loop dilution) will eventually achieve a
pure culture
When you see a slide for the 1st time you always put the objective on what setting?
scanning objective
the last objective is the oil immersion it magnifies 100 times for 1000 times with the eyepiece
this objective is the hardest to use. Oil immersion will be use to prevent the light from scattering and able to see the objective better.
Broth in Media inoculation technique is used
to grow large numbers quickly
Use a loop when ..
transferring to a broth
Compound Microscopes
use one magnifying lens and light and magnifies up to 1000 times
Antagonism
when one organism prevents other organisms from growing in its vacenaty
What is streak plating?
when you drag from one culture to another making a "z" looking motion.
How do you clean the lens?
with lens paper/cleaning fluid
the low power objective magnifies
10 times so 100 times with eyepiece
What technique is used to achieve isolation?
Streak plate