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Portage Learning / BIOD 171 microbiology Labs1-9

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TK01 Keeping a Lab Notebook Microbiology Lab1 Course Objectives: Cultivation of samples (growth conditions) - equipment used Identification of samples (biochemical assays) - tests available Evaluation of samples (microscopy) - visualizations and recognitions key concepts Basic Equipment: 1. Cleaning: autoclave 125 ° C, steamed, heat and pressure sterilization, chamber with thick door, not dry air but steamed - more efficient, make sure depressurized before opening Time restrictions: no autoclave then use steam (minutes) vs. hot air (hours) 2. Growing: fixed incubator = 37 ° C; shaker incubator = 37 ° C, for growth mediator, rotates and shakes in circular motions to aerate and help optimize bacterial growth 3. Visualizing: Microscopy, can't see with just the naked eye, must know parts of microscope 4. Storing: Refrigerate at 4 ° C, slows bacterial growth, prolongs life of sample at this temperature, when not finished with experimentation

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Portage Learning / BIOD 171 microbiology Labs1-9 2021

TK01 Keeping a Lab Notebook
Microbiology Lab1
Course Objectives:
Cultivation of samples (growth conditions) - equipment used
Identification of samples (biochemical assays) - tests available
Evaluation of samples (microscopy) - visualizations and recognitions key concepts


Basic Equipment:
1. Cleaning: autoclave 125 ° C, steamed, heat and pressure sterilization, chamber with thick
door, not dry air but steamed - more efficient, make sure depressurized before opening
Time restrictions: no autoclave then use steam (minutes) vs. hot air (hours)
2. Growing: fixed incubator = 37 ° C; shaker incubator = 37 ° C, for growth mediator, rotates and
shakes in circular motions to aerate and help optimize bacterial growth
3. Visualizing: Microscopy, can't see with just the naked eye, must know parts of microscope
4. Storing: Refrigerate at 4 ° C, slows bacterial growth, prolongs life of sample at this
temperature, when not finished with experimentation


Lab Safety:
1. Never eat or drink in lab - contamination risks
2. Wear Personal Protective Equipment (PPE):
Use gloves: latex, nitrile (hypoallergenic) purple,blue or green, cold thermal - liquid
nitrogen, etc., heat thermal resistant gloves - extreme hot conditions
Eyewear protection - goggles
Lab coat - dependent on working with materials that may be spilt on clothes
3. NEVER leave lab while wearing PPE includes: Public Hallway, Restroom, Cafeteria


Lab Notebook:
First Name Initial, Last name initial, Experiment #, Title of Experiment
Example: TK01 Keeping a Lab Notebook


Objective: Developing an organized lab notebook for documenting experiments, data,
procedures and records.

,Portage Learning / BIOD 171 microbiology Labs1-9 2021

Procedure:


1. Each step of lab protocol will be documented
2. Notate any clear deviations of lab experiment in red
Notes:
Additional information will be documented for future referencing.


Results:
Summary of final outcome of experiment, along with conclusions or the gist of the main
point from the experiment.



1. What three elements are used in an autoclave to sterilize equipment?


heat, pressure, and steam


2. What is the minimum temperature an autoclave must be set at to achieve sterile condition?


125°C


3. If you are working in a lab in which an autoclave is not available, and you are pressed for time, which would you chose to
best sterilize your equipment? Hot steam or hot air? Explain why you chose your answer.


Hot steam is the best choice as you can achieve a sterile environment in a matter of minutes whereas hot air will
take several hours to achieve the same effect.


4. What type of incubator is pictured below?


Fixed incubator

,Portage Learning / BIOD 171 microbiology Labs1-9 2021




Answer the following questions.



1. At what temperature is the fixed incubator set to, as presented in the lab video?


37°C


2. At what temperature should you refrigerate bacterial samples? Explain why this is beneficial.


4°C. This temperature slows bacterial growth and prolongs the life of the sample.


3. What are the FOUR types of gloves presented in the lab video?


Latex, Nitrile, Thermal cold, Thermal heat


4. What THREE rules were discussed in regards to lab safety that would protect you and others from contamination?


1. Never eat or drink in the lab


2. Always wear appropriate PPE


3. Never leave the lab wearing PPE


5. What are the main sections that should be found in a lab notebook? Name at least 4.


Objective, Procedure, Notes, Results and Deviations


Answer the following questions.

, Portage Learning / BIOD 171 microbiology Labs1-9 2021

1. You are a lab instructor and Paul Smith has turned in his lab notebook for you to grade the lab experiment #2 on
microscopy. Based upon what was covered in the lab video, how should Paul have entitled his lab experiment?


PS02 Microscopy


2. You arrive to your first day of work at a new lab. You are taking over for someone who took a new job at another lab. Your
boss informs you that because of time restraints, this person did not exactly follow the experimental protocol. In order to
proceed, you must know what he did differently. (1) According to the lab lecture, under what section of his notebook would
you look to find the experimental steps? (2) As changes to the experimental steps were made, what are these differences
called and how should it appear in the lab notebook?


(1) Procedure—this is where the steps for the experimental protocol are recorded.


(2) Deviations. All deviations should be written in red to immediately bring attention to the changes in the protocol.




TK02 Basics of Microscopy
Objectives:
Familiarity of the basic components of a light microscope and how to load a sample for viewing.


Procedures:
1. Review parts / components
2. Load sample slide onto microscope
3. Select magnification
4. Make necessary adjustments to optimize sample visualization
Notes:
Microscopy- makes objects larger for examination
Basic parts/components:
eyepiece - for viewing
arm / neck - grab microscope
objectives - 4x = shortest, 10x = yellow, 40x = dry objective,blue, 100x = oil objective, white
revolving nosepiece
stage - place the sample
stage controls

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