ROMANOWSKY STAIN AND DMITRI LEONIDOVICH ROMANOWSKY

Romanowsky is credited with the most important staining method used in hematology. 

Was he the first person or is there anyone else before him who tried a similar staining? 

Romanowsky introduced this staining in 1891.

Paul Ehrlich in 1870

Even before this, in 1870, Paul Ehrlich made a significant contribution of todays blood stain. He made a neutral stain which had a mixture of acidic dye (acid fuchsin) and basic dye (methylene blue) for blood smears.

To make it more interesting, I will tell you how he made this neutral stain. Ehrlich poured saturated aqueous solution of methylene blue into saturated aqueous acid fuchsin solution until the supernatant liquid was clear. Following this, he added more acid fuchsin until the precipitate redissolved. Methylene blue is said to be as one of Ehrlich favourite dyes.

So what colour this neutral stain gave to blood cells?

It stained the red blood cells and eosinophil granules as red, nuclei deep blue, lymphocyte cytoplasm lighter blue. This stain in addition also stained the previously unstained granules in the polymorphonuclear cells. His choice of using methylene blue as the basic component of neutral stain is an very important milestone in staining blood smears.

Cheslav Ivanovich Chenzinsky in 1888

Then, In 1888, Cheslav Ivanovich Chenzinsky who was a well known Polish pathologist used basic dye (methylene blue) and eosin instead of acid fuchsin (acidic dye) used by Ehrlich. This stain could stain the malarial parasite but not its nucleus.

Paul Ehrlich and Cheslav Chenzinsky

Both Ehrlich and Chenzinsky used acidic and basic dyes combination. Infact, they also used methylene blue and eosin and the same was used by Romanowsky.

Then what was the difference between these combinations? 

The difference is Romanowsky used aged methylene blue which gave the characteristic purple hue and this is the basis of Romanowsky effect.

Romanowsky published his preliminary findings of this blood stain where he used a combination of aged methylene blue and eosin in 1890. This used of polychromed methylene blue differentiated it from others.

So after his publications, the next query was how he modified this methylene blue which others dint think of doing? He published the answer for the same in 1891 in his doctoral thesis where he described he used methylene blue after mold began forming on the surface.

When I read this, question which came to my mind was,
Did Romanowsky really knew that polychromed methylene blue might produce this effect or was it a serendipity?

I could not get the answer for this, if you know or could find, please post in the comments section.

Till now we talked about three people, Ehrlich, Chenzinsky and Romanowsky and the list is not complete yet.

There is yet another person who also has independently observed the same stain combination as Romanowksy.

Who is he ???

We will see about him in the next post.

References:

1.     KP Krafts, E Hempelmann and BJ Oleksyn. The color purple: from royalty to laboratory, with apologies to Malachowski. Biotechnic & Histochemistry 2011, 86(1): 7–35.

2.    The picture of Ehrlich taken from google images, Wikipedia.org.

3.    The picture of Cheslav Chenzinsky is taken from KP Krafts, E Hempelmann and BJ Oleksyn. The color purple: from royalty to laboratory, with apologies to Malachowski. Biotechnic & Histochemistry 2011, 86(1): 7–35.

                                                             Written By Dr.Priyavadhana B