Gram Stain - Decolorizing

[tagz]

Anyone have tips to ensure that you are decolorizing the slide correctly? Here are two pictures of the same bacteria. I used less decolorizer on the second. There are some dark rods in the second sample; I'm just not sure if that's due to not using enough decolorizer in the second or too much in the first. Any help would be appreciated.

View attachment ImageUploadedByHome Brew1472753224.611615.jpg

View attachment ImageUploadedByHome Brew1472753244.994698.jpg

 

[BrewSRQ]

When I was doing them frequently, we would hold at a 45 and drip 95% ethanol until the runoff just turned clear. Time wise would be about ~5 seconds. This was easier to visualize than adding drops on a horizontal slide and waiting for a set yime.

 

[tagz]

That's the procedure I followed on the first. The second I used less and could see that not all the violet washed out before the rinse. So perhaps the dark rods on the second are just not decolorized properly and this would be a gram negative?

 

[BrewSRQ]

Yea, I'd say so.