Tissue staining

رنگ آمیزی عمومی

Tissue staining stage :

this step includes 2 types of general and specific methods . the general methods is : Hematoxylin-eosin method , with this method the nucleus turns to purple and cytoplasm turns to pink.

The specific method is that each cell component takes a specific color, which includes staining of frozen sections , connective tissue , neural system , staining for some cytoplasmic materials , staining for enzymes , staining for micro organisms and cytological staining.

Solutions used in routine staining include : Ethanol, xylene, Hematoxylin , Eosin , Hydrochloric Acid, lithium Carbonate, Distilled water, Acetic Acid.


0.5 cc of Acetic Acid should be added to every 100 cc of Hematoxyline before use.


Place the sections in the oven at temperature of 120◦ C for 15-20 minutes before staining , letting them stick completely to the slides.

  1. pure xylene (1-2 minutes)
  2. pure xylene (1-2 minutes)
  3. pure xylene (1-2 minutes)
  4.  ethanol 96% (2-3 minutes)
  5.  ethanol 80% (2-3 minutes)
  6.  ethanol 70% (5 minutes)

Rinsed with distilled water( temperature about 36 degree)

7.  Hematoxylins : 5 to 10 minutes, red

Rinsed with distilled water

8. 1% alcohol acid (prepare 70% alcohol and pour 1 cc of concentrated hydrochloric acid for every 100 cc of alcohol)

9. 1% saturated lithium carbonate: several dips (per 100cc of distilled water, one gram of lithium carbonate powder, blue)

10. Eosin (4 to 6 minutes)

Rinsed with distilled water

11. 70% alcohol (1-2 minutes)120

12. alcohol 80% (1-2 minutes)

13. alcohol 96% (1-2 minutes)

14. alcohol 96% (1-2 minutes)

15. mixture of alcohol and xylene (1 volume alcohol 96%, 3 volume xylene)

16. pure xylene (1-2 minutes)

17.pure xylene (1-2 minutes)

18.pure xylene (1-2 minutes)

رنگ آمیزی بافت

Figure1: an example of Kidney Tissue stained by H&E method


This method of staining strongly highlight the nucleus of the cells that have turned to blue color, & also their morphology & location.


Figure2: detection of nucleus , location & morphology of cells by H&E method

It is also possible to observe the density of the cells in a specific area & the divided cells by H&E staining under a microscope.

رنگ آمیزی- بافت

Figure 3: identification of divided cells & cells density by H&E method.

Using this method it is possible to size the cells  & observe their structure and that is because all tissues contain blood vessels & this leads to the possibility of comparing the size of the tissue cells.

tissue staining
tissue staining

Figure 4: estimating the size of the cells by H&E staining method

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