Day 2 In the Journey to New Discoveries

May 15, 2014

Being that we had a quiz in lecture, all of us were feeling a bit tired and overwhelmed from learning, what we felt was, everything a person could possibly know about viruses. Regardless, we were excited and curious to see what growth our unknown bacteria underwent.

Part 1 of Today's Lab: Observing Growth

After being instructed to remove our sample bacteria slants, plate, and broth from the 37 degree Celsius incubator, we sat down to examine the changes they had undergone. With simply a naked eye, it was obvious the bacteria had grown.

1. The Plate
   The plate had the most obvious growth of bacteria out of our three different bacteria samples. Under microscopic view, we were able to see that multiple single colonies had emerged along the boarder. According to Dr. P, our bacteria was “a fast growing one.” The visible characteristics of the bacteria are as follows:
   

            Colony Pigmentation: cream/tan

   
   
   Whole Colony (form): circular
   
   Colony Margin (edge): undulate
   
   Elevation of Colony: hilly
   
   

                 II.The Broth

While it wasn't obvious at first glance that the bacteria had grown overnight, we performed a flicking motion against the bottom of the test tube to stir up the bacteria sediment. The broth then became more cloudy/hazy.

                 III.The Agar Slant

 Observing the bacteria growth both with the naked eye and under the microscope, we were able to deduce that the bacteria growth on the agar slant was beaded.

Part 2 of today's lab: Observing the Motility of Our Unknown Bacteria

Being that our bacteria had growth, we then prepared, still tired but certainly interested, to see if our bacteria was alive and moving.

Materials used:

Cover Piece

Bunsen Burner

Loop Tool

Depression Slide

Microscope 

Unknown Bacteria

Vaseline 

Procedure:

   Using a loop and of course, aseptic technique, we took a sample of bacteria from our broth and after flicking the bottom to ensure a well mixed sample, we placed a drop on a cover piece provided. After adding vaseline to the corners, we attached it to the depression slide and flipped it over. This type of experiment is known as the Hanging Drop Experiment.

With a compound microscope, we decreased the light completely to ensure visibility of the bacteria. Between our compound microscope and Dr. Pathakamuri's microscope, we were able to see that our bacteria was swimming around.

Dr. P suggested that we add broth to the cover slide to further observe the bacteria's motility. The projected image of the microscope's view revealed that the bacteria seemed to “stand up” in attempts of broth consumption.

Part 3 (our favorite part) of today's lab: Simple Staining

Following our observation of the bacteria's motility, we were instructed by Dr. P to begin a simple stain experiment on the bacteria.

Materials Used:

Slide

Loop

Bunsen Burner

Crystal Violet dye

DI H2O

Tongs

Microscope

Bibulous Paper

Procedure:

We began by cleaning the slide with a task wipe and, practicing aseptic technique throughout, we obtained a sample of our bacteria from our plate and spread it into a thin film on the slide. We allowed it to air dry in order that the moisture in the sample evaporate. Then, using tongs, we swiftly passed in through a flame, bacteria side facing up, three times. The bacteria was then considered “heat-fixed.”

 We covered the fixed smear with several drops of crystal violet stain and let is sit for 25 seconds, as directed by the lab manual. Then, we rinsed the slide with DI H2O to remove excess stain and blotted the water with pieces of bibulous paper.

After that, we examined the newly stained bacteria under a blue light to discover the shape of the bacteria.

Under the lowest magnification, it appeared that our bacteria was round. However, under the highest magnification, we discovered that our group's bacteria had groups of small rods.

This concluded day 2. We look forward to seeing new growth tomorrow and experimenting to discover our bacteria's identity .

Joanna Dawyot, Cassie Livingston, Mary Rose Capara 

DisclaimerAll content provided on this blog is representation of the blog owner and not FranciscanUniversity of Steubenville. The information on this site is purely used for education purpose. The owner of this blog makes no representations as to the accuracy or completeness of any information on this site or found by following any link on this site. The owner will not be liable for any errors or omissions in this information nor for the availability of this information. The owner will not be liable for any losses, injuries, or damages from the display or use of this information. Privacy The owner of this blog does not share personal information with third-parties nor does the owner store information is collected about your visit for use other than to analyze content performance through the use of cookies, which you can turn off at anytime by modifying your Internet

 browser’s settings. The owner is not responsible for the republishing of the content found on this

 blog on other Web sites or media without permission.Blog CommentsThe owner of this blog reserves the right to edit or delete any comments submitted to this blog without notice due to;1. Comments deemed to be spam or questionable spam2. Comments including profanity3. Comments containing language or concepts that could be deemed offensive4. Comments that attack a person individuallyThis policy is subject to change at anytime.