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Gram staining

 
 
Vivace
 
Sat 24 Oct, 2015 01:35 pm
in an Gram staining experiment of E. coli, one student observed a bright pink staining while the another student only observed faint staining. Why is this happening?
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Type: Question • Score: 2 • Views: 1,829 • Replies: 5
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farmerman
 
  2  
Sat 24 Oct, 2015 02:25 pm
@Vivace,
did you each do your own staining procedures? If so, maybe the one who observed the faint staining left the ethanol go too long
Vivace
 
  1  
Sat 24 Oct, 2015 09:22 pm
@farmerman,
Yup each doing their own staining. But safranin is added after the ethanol decolourization. How would that affects the Counterstain?
farmerman
 
  2  
Sun 25 Oct, 2015 06:07 am
@Vivace,
safranin is one of a family of stains that , after decolorization, will keep the gram negatives pink and the positives purple (or is it the orther way round, its been so damn long)

prep
crystal violet
ethanol decolorization
wash
add counterstain
My point was that. maybe the ethanol wash was done to vigorously (too long) and that weakened the counterstains affinity.


ossobucco used to be a lab microbiologist so shed know
Vivace
 
  1  
Sun 25 Oct, 2015 07:12 am
@farmerman,
Alright. Thanks.
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ossobuco
 
  1  
Sun 25 Oct, 2015 10:53 am
@farmerman,
No, I don't know.
Though my degree is in bacti, I last ran the bacti part of a lab in 1967.. most of my med lab years were in immunology/rheumatology/hematology; I stopped working in the medical field in '80...

but what you are saying makes sense to me.

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