Question

What are some ways in which a Gram stain procedure can go wrong? How would you troubleshoot those problems?

Answer 1

Decolorization step is an important part in it,when over decolorization will lead to loss of primary stain making gram positive bacteria To appear gram negative. So to avoid this proper quantity of the decolorizing agent to be added

When the smear is thicker ,the dye cannot evenly penetrate the entire surface causing wrong results. So the smear should be thin and even to get the accurate results

When the bacterial culture is old with more than 18hours incubation can interfere result by making gram positive bacteria to look pink

Bacteria grown on EMB can give variable staining report

When the gram staining kit is out dated

In the first staining step if gram stain is done in heating procedure then all the mycobacterium can appear is gram positive

Improper storage can interfer with the results.