Student's Name: ______________________________ Date:____________

Instructor's Name:______________________________ Score:____________

Course No. and Course Code:_____________________

LABORATORY SAFETY AND BASIC LABORATORY TECHNIQUES

Activity No. 1

OBJECTIVES:
1. _______________________________________________________
2. _______________________________________________________

Safety in the laboratory is of the utmost importance when doing practical work in the laboratory.
The following guidelines should be observed to ensure safety in the laboratory/
1. Do not drink nor eat in the laboratory.
2. Wear your Personal Protective Equipment (PPE) at all times during the conduct of the
laboratory exercises. Store laboratory gowns properly in the lockers. Never bring them home
except for laundry purposes. Separate your laboratory gowns from the rest of the clothes and
soak it with household bleach to disinfect it prior to washing.
3. Wear appropriate footwear. Closed toe shoes are recommended. Wearing of sandals or open
toed footwear is prohibited.
4. Use of electronic devices in the laboratory (ex. Cellphones, tablets, etc.) are highly
discouraged, except when documentation of results/laboratory demonstrations necessitates the
use of such device.
5. Bags, books and other personal belongings should be placed at a designated area near the
laboratory. They should never be placed on bench tops that serve as your working area.
6. Behave in a responsible manner. Horseplay, practical jokes and pranks are not tolerated.
7. Female students with long hair should tie back their hair or enclose it in a hair net. This
prevents possible fire hazards when working with alcohol lamps and burners.
8. Never pipet by mouth. Use appropriate mechanical pipetting device.
9. Never attempt to light an alcohol lamp with another alcohol lamp. This presents a possible fire
hazards.
10. Avoid touching any part of your face with your gloves. This exposes you to probable
contamination.
11. Use test tube racks or trays to transport tubes or any glasswares in the laboratory. This limits
the probability of breakage and further laboratory accidents.
12. Do not take out or transport any bacterial culture out of the laboratory premises.
13. Report any spillage or breakage of glassware to your laboratory preceptors. A spill kit should
be on hand to take care of any spillage/breakages.
14. Immediately report any laboratory accident/s involving your classmates to your laboratory
preceptors for immediate action (ex. Giving first aid to burn victims)
15. Disinfect your work area with any appropriate disinfectant available prior to conducting the
laboratory experiment. Do likewise at the end of each laboratory session.

BASIC LABORATORY

Working in a microbiology laboratory or any other laboratory for that matter, requires the
individual to be at least familiar with the basics. One must always remember that live organisms
are handled almost every lab session. It is very important to conduct a risk assessment before any
laboratory activity commences. Safety guideline must be in place and protocols for laboratory
accidents such as spillage, proper decontamination procedures, breakage of glass wares, etc. be
established. In risk assessment, the laboratory instructor is mainly tasked with assessing several
factors such as:
a. Level of competency of staff
b. Degree of risk with organism/s being handled in class
c. Suitability of laboratory facilities- example: presence of incubators, autoclave, Bunsen
burners/alcohol lamp, racks to hold cultures, laboratory tables, sink, etc.
d. Source of cultures- if the cultures are clinical isolates or control organisms from reputable
sources
e. Proper disposal procedures- presence of autoclave and containers with disinfectant for discard

Good Microbiology Laboratory Practices must be observed at all times to prevent contamination
of the laboratory activity being conducted as well as causing probable contamination/infection of
others. Aseptic techniques should be practiced at all times. Decontamination of work surfaces,
before and after any laboratory activity must be encouraged.
Basic tools used in a Microbiology Laboratory consist of the following:
1. Disposable wire loops and needles
2. Reusable inoculating loops
3. Alcohol lamp/Bunsen burners
4. Disposable Pasteur pipettes
5. Petri dish
6. Test tube rack
7. Test tubes/glasswares
8. Hot plate/stirrer
9. Serological pipette/pipette aid
10. Weighing scales (top-loading balance/analytical balance)
Instructions on the proper usage of these basic tools is the responsibility of the Laboratory
Instructor

Improper use may lead to accidents and damage to the equipment. Aseptic technique is a
procedure done in the laboratory to ensure freedom from potential pathogens, preventing
contamination and creating a safe working environment. It is a skill that is developed and is
practically done at every laboratory activity.
Practical pointers before embarking on a “hands-on” laboratory activity:
1. Decontaminate work surfaces with an appropriate disinfectant (example: 70% alcohol, Lysol)
2. Make sure that you have everything you need for the laboratory exercise within easy reach
3. Do not forget to wear appropriate Personal Protective Equipment (PPE). Example: laboratory
gowns, masks, gloves, eye shields (optional)
4. A container for discard with an appropriate disinfectant should be present

Aseptic Techniques
Flame sterilization of inoculating loops
Below is an illustration of how to properly flame sterilize an inoculating loop.
Transfer of Broth Culture

When transferring culture from a broth in a tube, make sure that you shake the tube from side to
side or by tapping the tube while holding it firmly with your hand. This is to ensure that there is
homogenous suspension of the bacteria. Heat the loop and entire wire red hot. Allow to cool.
Remove the cap or cotton plug with your little finger and flame the tip of the tube. Insert the loop
and carefully withdraw the loop with the organism. Heat the tip of the tube and return the
cap/cotton plug.

Inoculation of Plates

Plates to be inoculated with any organism/specimen should be completely dry before use. It is
recommended that the plates be placed in the incubator prior to the experiment. This is to prevent
the buildup of moisture that could contribute to contamination as well as to ensure that isolated
colonies will grow on the plate. Check the plates for possible growth. Discard plates with any
growth. Flame sterilize loop before use. Allow to cool. Set aside. Make sure it does not come
into contact with anything after it has been flame sterilized. With your free hand, open the lid by
raising it just enough to gain access to the agar to be inoculated. Do not remove the cover and
place it on top of the table, leaving the media exposed to possible external contamination.
Transfer loopful of organism from the broth culture and streak for isolation. Label plates
accordingly and include the date of inoculation. Incubate plates in an inverted position, bottom
side up.

Check the agar slants for possible contamination. Discard any slant with growth. Observe aseptic
technique. Transfer organism into the slants by removing the cao/cotton plug with your little
finger, and flame the tip of the tube. Inoculate slant with the loopful of organism by spreading it
evenly across the entire agar slant. Label slants accordingly and include the date of inoculation.

Reminder:
1. Decontaminate working area after the laboratory exercise.
2. All manner of discard: plates, tubes, etc. should be autoclaved first before washing and discard
treated as ordinary waste.
3. Dispose of gloves, mask in the proper waste bins.
4. Leave your laboratory gowns in your lockers; do not bring them home.
5. Wash hands thoroughly before leaving the laboratory.

CONCLUSION: