BLOOD MADE COLORFUL: ROMANOWSKY STAINS

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Romanowsky staining is the most important stain used in hematology and cytology.

It is a differential stain.

Stains based on the principle of romanowsky principle are Leishman, Giemsa, Jenner, Wright, May- Grunwald stain and Field Stains.

This staining technique is named after the Russian physician Dmitri Leonidovich Romanowsky who developed this stain in 1891. The original stain used were eosin Y and aged solutions of methylene blue.


                         Dmitri Leonidovich Romanowsky (Picture taken from Google Images, Wikipedia.org)

Principle: The stain consists of both cationic dye and anionic dyes. The cationic dye used is Azure B and the anionic dye is eosin Y in aqueous solutions. These two dyes act separately or in conjunction on pre-treated biological substrates to give a typical staining pattern which depends on the chemical and physical properties of the substrate.

There is some concept known as Romanowsky effect.

I  was not much clear on this concept during my initial MD days.

The following definition of Romanowsky effect is from British Journal of Hematology

The unique effect of Romanowsky staining is based on the fact that with biological substrates the blue cationic dye Azure B and the red-orange dye eosin Y give more colours than just blue and red-orange. Purple is the most important colour which characterizes the Romanowsky effect. Satisfactory stains should give the purple colour in cell nuclei and this provides strong contrast against the colour blue which is typical for cytoplasm rich in RNA.

The colours which were formed were unattributable to these staining components alone and these distinctive shades made us to recognize different cell types.

In some stains, eosin Y is combined with polychrome methylene blue instead of pure Azure B or sometimes mixture of catioinic thiazine dyes are used.

Was Romanowksy the first person to try this staining?
Leave your opinions in the comments section.

See you all in the next post with the answer.

References:

1.     ICSH reference method for staining of blood and bone marrow films by azure B and eosin Y (Romanowsky stain). International Committee for Standardization in Haematology.  British Journal of Haematology. 1984:57;707-710.
    
                                                                                                             Written By Dr.Priyavadhana


Comments

  1. bpranjan18 July 2020 at 23:23

    Interesting blog I read so far.. We didnt know the story behind the stain.. came to know from your blog. Keep doing more like thiss..

    ReplyDelete
  2. bpranjan18 July 2020 at 23:24

    In 1891 itself he identified this stain.. So i think he s the one who tried 1st

    ReplyDelete
    Replies
    1. Priyavadhana19 July 2020 at 13:05

      Ok Ranjan.. We shall discuss in detail in future posts.

      Delete
  3. Unknown19 July 2020 at 10:39

    Nice ma'am

    ReplyDelete
  4. Dr Deepak Verma19 July 2020 at 20:08

    NICE VERY INFORMATIVE.

    ReplyDelete

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