THE AMERICAN 



MONTHLY 



MICROSCOPICAL JOURNAL. 



Vol. VI. 



Washington, D. C, March, 1885. 



No. 3. 



Stjiining Tissues for Photou:raph.v. 



BV GEORGE A. PIERSOL, M. D. 



Satisfiictorv results in photograph- 

 ing histological tissues depend largely 

 upon two conditions : — 



1. Having a section so thin and 

 even that little more than a single 

 laver of cells is included ; 



2. Having such thin section pro- 

 perly stained — especially sufficiently 

 difterentiated. 



Regarding the first condition, but 

 little difficulty is experienced in these 

 days of sliding microtomes, v^hose 

 advent has marked a new^ era in sec- 

 tion cutting. 



The successfvd completion of the 

 second condition for photography, is 

 not always as readily accomplished. 

 Bv most workers, probably, the stains 

 ordinarily employed and valued for 

 general use are borax-carmine and 

 hematoxylin ; of the two, the latter 

 is usually the more highly prized — 

 the simple manipulations required 

 and the unsurpassed results justly 

 giving hematoxylin a recognized pre- 

 eminence. 



These sections stained with boi'ax- 

 carmine (properly used) yield often 

 excellent negatives ; in their strata 

 the red color being sufficiently non- 

 actinic to give a vigorous contrast on 

 the plate. In well differentiated car- 

 mine staining, however, little else 

 than cells is colored, and frequently 

 delicate detailsoftheconnective-tissues 

 are wanting on account of their trans- 

 parency. 



Hematoxylin stainings, in very thin 

 sections, while all that can be desired 

 under the microscope, are usually very 

 disappointing when photographed ; 



the delicate layer of tissue offers al- 

 most no actinic contrast when mono- 

 chromatic sun-light is obtained by the 

 ammonio-sulphate of copper cell. 



Since hematoxylin is so extensively 

 cmplo*yed in all lines of work, a ready 

 modification of this staining to meet 

 the needs of photography is of advan- 

 tage. Such a result is obtained by a 

 modified use of a formula of Wiegert, 

 already commended to the readers of 

 the Journal by Dr. Councilman* 

 for the study of the brain and spinal 

 cord. While especially intended for 

 nervous tissues, the modified use fur- 

 nishes specimens of all organs ad- 

 mirably adapted for photography. 



No especial formula for hematoxy- 

 lin is needed, using one which is ca- 

 pable of staining deeply and giving 

 standard results. In the usual course 

 of work the sections are stained; a 

 few very thin ones, however, are al- 

 lowed to remain in the solution, after 

 those for ordinary preparation, until 

 they are of an intense dark purple, 

 when they are transferred, one by 

 one, to a capsule containing a solu- 

 tion composed of the following con- 

 stituents : — 



Borax i • 



Potassium ferricyanide 2.5 



Water 100. 



In this they are kept moving until 

 the intense color is gradually dis- 

 charged, and the purple tint is re- 

 placed by a bronze-yellow, shading 

 to saffron. Before the sections reach 

 the latter color they should be washed 

 in water ; the further usual steps in 

 mounting are then completed. 



*The Microscopic Investigation of the Brain and 

 Spinal Cord, vol. v, p. 201. 



