Advances in fluorescent dyes and instrumentation were crucial in the development of the fluorescence microscope. 

First, I will present some examples from the plethora of organic dyes and stains that were used in the fluorescence microscopy of biological specimens.

 

The development of the synthetic dye industry by William Perkin (1838–1907) in 1856 resulted in the synthesis and the chemical and spectral characterization of many new fluorescent dyes.

 

Acridine orange was synthesized by Bender in 1989; acriflavine (a mixture of dyes) was synthesized by Erlich and Beneder in 1912; both dyes were used in histology. Auramine O was synthesized by Kern and Caro in 1883; it was used to stain bacteria. Caro synthesized Eosin B in 1875; it was used as a counterstain.

 

Adolph Von Baeyer (1871) a German chemist, synthesized Spiro[isobenzofuran-1(3H),9'-[9H]xanthen]-3-one, 3',6'-dihydroxy.

He apparently coined the name “fluorescein”, from “fluo” and resorcin, (resorcinol) which he reacted with phthalic anhydride. In 1905 he was awarded the Nobel Prize in Chemistry "in recognition of his services in the advancement of organic chemistry and the chemical industry, through his work on organic dyes and hydroaromatic compounds".

 

The dye fluorescein was first synthesized (Adolph Von Baeyer) in 1871 and it has a long history connected with cell staining.

 

In 1881, the bacteriologist Paul Ehrlich (1854–1915) used fluorescein to observe the passage of aqueous humour in the eye. He also used various aniline fluorescent dyes to stain bacteria and thereby increase their contrast in the microscope. Rhodamine B and Rhodamine G was synthesized by Ceresole in 1887 and 1891, respectively, both dyes were used as vital fluorochromes to stain living tissues. 

 

Before the 1911–1913 invention of the fluorescence microscope there were many stains and dyes for use in histology and cell biology (Kasten, 1989). Then with the invention and commercial availability of the fluorescence microscope many cells and tissues were labelled with dyes and their fluorescence was studied. 

 

 

Fluorescein videos: