Lab 1 - Basic Laboratory Techniques

Be Sure to Know Where These Are:

• The location of the safety shower

• The eye wash station

• The fire extinguisher

• The first aid kit

When Entering the Lab

1) When you first enter the lab, place your backpack, coat and anything else you are carrying under the lab bench.

2) Choose a lab cost from the costs hanging on either side of the microscope shelves.

3) Then wash your hands using antibacterial soap.

4) Go to your place and wipe down the bench using PREempt.

When Exciting the Lab

1) When you have completed your work, wipe down the bench again with PREempt.

2) Hang up your lab coat

3) Pick up your stuff and leave

Purpose of Aseptic Technique

• Protect your work from contamination

• Protect yourself from the bacteria you are working with

• Protect the people you come in contact with from the bacteria you are working with.

Lab Containment Level 1 Minimun Requirements

A. Must wear a lab coat in the lab at all times.

B. Must wear safety glasses when working with liquid cultures.

C. Cannot eat or drink in the lab.

D. Any open cuts or scrapes must be covered with a waterproof dressing.

E. Only necessary items on the lab bench.

F. Long hair must be tied back.

G. No open-toe shoes.

H. No contact lenses.

I. No elaborate rings or dangly jewellery.

Cell Phone is the lab

• Need to be disinfected when entering the lab and disinfected again just prior to leaving the lab

• Put 70% ethanol on a paper towel or kimwipe and wipe down the phone

• While in the lab, the cell phone must be kept in a plastic bag in your lab coat pocket.

Four specified disposal sites for waste

• Two types of sharps containers

  • Only glass sharps

  • Only metal sharps

• Solid waste buckets

• Plate waste buckets

• Tube waste

Two types of sharps containers

All sharp objects, including microscope slides and other glassware, are to be disposed of in the sharps containers provided on each bench.

• Only glass sharps

• Only metal sharps

Only glass sharps

Go in the glass sharps container

Only metal sharps

Go in the metal sharps container.

Solid Waste Buckets

• All contaminated swabs, contaminated pipette wrappers and any paper towels used to clean up bacterial spills

• NOT paper towels used to dry hands or to disinfect the bench, instead, put these paper towels in the regular orange garbage bin.

Plate Waste Buckets

All petri plates are to be disposed of in the buckets labelled “PLATE WASTE”.

Tube Waste

• All tubes are to be placed in the racks on the side bench underneath the sign labelled “TUBE WASTE”. The tubes should be sorted according to size.

To properly mark it must include:

• Name and bench, if applicable

• Date it was inoculated

• Name of the bacteria or exercise

Once finished with observations

• Dispose of all media in the proper waste disposal sites

If you spill the bacterial culture

1) Tell Instructure, and it will help you.

2) If there is no one, follow the lab manual. Use first ethanol and then PREempt.

Oxygen Requirements (organisms)

• Obligate aerobes

• Obligate anaerobes

• Facultative anaerobes

• Aerotolerant anaerobes

• Microaerophiles

Obligate Aerobes

• This includes the organisms that need oxygen to grow/survive.

• See all the material toward the top of the media.

Obligate Anaerobes

• This includes the organisms that do NOT need oxygen to grow.

• Appear only at the bottom of the media.

Facultative Anaerobes

• Organisms can grow in the presence or absence of oxygen.

• In this situation, a bacterial culture will appear most abundant at the top of the media, then gradually decrease as it moves downward.

Aerotolerant Anaerobes

• They can live in the absence of oxygen (they can survive in its presence but do NOT use it).

• In a tube, a bacterium will have a uniform abundance throughout the media.

Parafilm

It is used to seal the plates if the bacteria on the plate are unknown.

Potential Results (Both characteristisc)

• Pellicle

• Flocculant

• Turbid

• Sediment

Pellicle (Broth Characteristics)

• Thick growth at the top of the media

Flocculant (Broth characteristics)

• Flaky aggregates disperaed throughout tube

Turbid (Broth characteristics)

• Growth throughout the tube

Sediment (Broth characteristics)

• Growth at the bottom of the tube.

Potential Results for Bacterial Growth on Slants

• Filiform

• Arborescent

• Beaded

• Effuse

• Rhizoid

• Echinulate

Filiform (Growth on slants)

• Following the line of the original streak.

Arborescent (Growth on slants)

• Treelike growth

Beaded (Growth on slants)

• Nonconfluent to semiconfluent colonies.

Effuse/Spreading (Growth on slant)

• Thin spreading growth

Rhizoid (Growth on slant)

• Rootlike growth

(Smaller branching then arborescent)

Echinulate (Growth on slant)

• Slight pointed spreading from the original line.

Bacterial Growth in Plates (factors)

• Shape

• Margin

• Elevation

• Size

• Texture

• Appearance

• Pigmentation

• Optical property

Shape (Bacterial growth on plates)

• Circular

• Rhizoid (rootlike)

• Irregular

• Filamentous (like paint)

• Spindle (oval)

Margin (Bacterial Growth on Plates)

• Entire

• Undulate

• Lobate

• Curled

• Rhizoid

• Filamentous

Elevation (Bacterial growth on plates)

• Flate

• Raised

• Convex

• Pulvinate

• Umbonate

Size (bacterial growth on plates)

• Punctiform

• Small

• Moderate

• large

Texture (bacterial growth on plates)

• Smooth

• Rough

Appearance (Bacterial growth on plates)

• Glistening (shiny)

• Dull

Pigmentation (Bacterial growth on plates)

• Nonpigmented (eg, cream, tan, white)

• Pigmented (eg, purple, red, yellow)

Optical Property (Bacterial growth on plates)

• Opaque

• Translucent

• Transparent