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Thursday, October 17, 2013

Endospore Stain

Next we wanted to see if our bacteria possessed any endospores. An endospore is a mechanism used by bacteria that is specialized for the survival of that bacteria in unfavorable conditions.

We began this staining procedure much like we did for the Acid Fast stain. First we smeared some of our sample bacteria on the the slide and using a Bunsen burner we heat fixed the bacteria to the slide so that it can we stained and viewed under a microscope.
After that we put a piece of bibulous paper over the slide and using forceps, moved the slide onto a drying rack over a beaker of boiling water.


Next we saturated the paper with malachite green solution. Just like the acid fast stain we had to remain attentive and continue to add drops so that the solution did not evaporate and dry out the slide. 
We continued this practice for 6 minutes, constantly adding more drops because the solution was evaporating very quickly from the bibulous paper. 


Using forceps we removed the bibulous paper from the slide and placed it in a bio-hazard bag. We then transported our slide back to our lab bench, using forceps, and placed it on a drying rack over the sink in order to let it cool off. We then rinsed the slide off with water to remove excess malachite green solution. 




 Next we covered the smear with safranin for 90 seconds. Then we washed off the excess safranin stain with water.  


To completely dry the stain we blotted the side between bibulous paper, being careful to not blot it too excessively.

Now our slide is ready to view under the oil immersion lens of our microscope. Upon viewing our bacteria we were able to conclude that our bacteria does not possess endospores. If our bacteria did we would be able to see little green dots at the ends of our bacteria rods. But our bacteria is completely red, so they do not have endospores. 



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