1887.] MICROSCOPICAL JOURNAL. 83 
No. e78: —Green gland of cray-fish—killed in atmosphere of chloroform. March 15, 1887. 
Mo. Day. Hour. Mo. Day. Hour. 
Corrosive sublimate, saturated Washed and transferred to 100 
aqueous sol., temp. 18° per cent. alcohol............... 4 Il 10,25A.M. 
BEM iets son eee ccctamditecessavena 3 15 2.00 P. M. @hloroformvessssssscesssssceseeveeesens 4 II 12.20P.M. 
EAPO (GSES) roe-cenes:-2esscceesiecersse 3 15 Bf Ly and paraffine........ 4 II Zigere 
Eopper cent. alcohol.:........---s:-- 3 15 {liters Pure paraffine at 55° cent........ 4 II QegOmmer 
ORPCISCENLE ees)  atscccesscesc cate B 15 fuer, S40 imibed dedPerecesseserescsateeeetteere 4 Ty TOR20n ce 
SOLeUX] CALMING)... decesc-c0ss loscsese 4 10 8.48 A.M. Mounted inchloroform balsam, 4 I2 2.00 A: M. 
Cut with Jung microtome. Ribbons cemented with collcdion. Cleared with turpentine. 
The following notes will be found helpful to any one who follows the fore- 
going course :— 
t. The saturated corrosive sublimate solution is made by heating water to 
boiling, and adding as much of the salt as will dissolve, then allowing the 
solution to cool, whereupon the excess of the salt will be crystalized out of 
the strong solution. 
2. The work of the corrosive solution is to coagulate the protoplasm of 
the cells. Since it acts rapidly, it soon hardens the peripheral parts of the 
piece, and then the ready attack upon the central cells is prevented. One 
should, therefore, never harden large pieces with corrosive sublimate, as 
cells separated from the fluid by more than | of an inch of intervening tissue 
will not be readily reached by the reagent, and so not well preserved. If it 
is desired to hurry the action, one may add a few drops of glacial acetic acid 
to the corrosive sublimate, and then leave the tissue in the fluid only ten to 
twenty minutes. 
3. After coagulation of the protoplasm by corrosive sublimate, the salt 
must be entirely removed from the tissue, and since warm water would tend 
to destroy the cells a steady stream of cold water flowing over the specimen” 
is employed. This must continue long enough to entirely remove the cor- 
rosive, viz., from one-half hour to one hour, or even more. If the water is 
cold it will not hurt the specimen. 
4. As soon as all the corrosive sublimate has been removed, the specimen 
must be gotten as soon as possible into strong alcohol. It must not, how- 
ever, be transferred directly from the water to 95 per cent. alcohol, but car- 
ried up through weaker alcohols, 30, 50, and 7o per cent. are a good series, 
and since a change in protoplasm will not go on in 7o per cent. alcohol the 
specimen may remain in it till it is desired to stain, precaution being taken 
to keep the alcohol at 70 per cent. 
5. Borax-carmine solution is made by dissolving 4 grs. borax and 4 grs. 
carmine in 100 cc. distilled water with heat, adding 100 cc. of 70 per cent. 
alcohol, and after 36 hours using filtered solution. The solution keeps with- 
out change, indefinitely, and may. be used over and over again: In using the 
borax-carmine, one immerses the whole piece (called staining ‘‘ zz ¢oto,” or 
‘¢in bulk,” or ‘‘in piece”). In this case the time was twenty-four hours, or 
thereabouts ; a longer time will do no harm. After removal from the borax- 
carmine, the piece will have a deep maroon color; this is to be changed to 
a bright red by washing a few moments in 7o per cent. alcohol, to which 
two parts of strong hydrochloric acid have been added. During this wash 
much of the staining fluid will be at first removed, but gradually less and 
less. It should be continued with perhaps a change of the solution until no 
more color is removed. 
6. The alcohol which is used to remove the last trace of water before the 
transfer of the piece to chloroform, the solvent of paraffin used in the present 
instance, need not be strictly 100 per cent., though where one is working 
with small quantities it is wiser to buy the absolute and be on the safe side ; 
but an alcohol of 97 per cent., or even 96 + per cent., will answer equally 
well if procurable. Druggists do not often dispense an alcohol of higher 
