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The page below is a sample from the LabCE course Troubleshooting Guidance for Hematoxylin and Eosin (H&E) Stain. Access the complete course and earn ASCLS P.A.C.E.-approved continuing education credits by subscribing online.

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This section of lymph node illustrates how a highly cellular sample
can give the impression of overstaining with hematoxylin.

This section of kidney shows a nice balance,
of hematoxylin and eosin.

Cellularity Makes a Difference

Highly cellular samples (eg, tonsil, lymph node) can be very concerning. Remember that lymphocytes have little cytoplasm and there is not nearly the cellular material between cells as with other tissues. For this reason, the hematoxylin does not have to compete with the eosin. The compact nature of the cells also concentrates the DNA, giving these highly cellular tissues the appearance of being overstained, when in reality, they may simply need to be sectioned thinner.
Note the difference between the gross staining intensity of the lymph node and the kidney. The high cellularity of the lymph node gives the appearance of an overstained slide.