ZOOLOGY AND BOTANY, MICROSCOPY, ETC. 
701 
liquid will now easily penetrate the shell and the mantle cavity. The 
chief stain used was alum-carmine, used to stain in bulk. It is con- 
venient to remove the foot and other organs with a razor or a pair of 
scissors, as a great deal of time and trouble may be saved, and the stains 
penetrate more easily. The sections were cut with Yung’s microtome, 
after imbedding in chloroform and paraffin. At Prof. Lankester’s 
suggestion use was also made of the injection method, though the author 
is strongly prejudiced against it, as he thinks it very likely to mislead. 
He used soluble Berlin blue, and injected by blowing the injection 
through a fine glass pipette with the mouth ; these injections confirmed 
the results obtained by dissections and sections. But the use of 
syringe and strong pressure resulted in the breaking of the walls at 
various points. 
Preparation of Nudibranchs.* — Most of the Nudibranchs whose 
cerata have been studied by Prof. W. A. Herdman and Mr. J. A. Clubb 
were killed and fixed with Kleinenberg’s picric acid, stained with picro- 
carmine, passed through graduated alcohols, imbedded in paraffin, and 
cut with the Cambridge rocking microtome. The specimens of Hermsea 
dendritica were obtained at Plymouth by Mr. Garstang, who plunged 
them for a moment, while alive, into glacial acetic acid, transferred them 
to a saturated solution of corrosive sublimate for half an hour or so, and 
then passed them through grades of alcohol. 
Study of Development of Limulus longispina.j — Mr. Kamakichi 
Kishinouye fixed the eggs of L. longispina by heating in water to 60° or 
70° C., or by plunging into water of that temperature. After cooling 
they were transferred to 70 per cent, alcohol, where they were left for 
one or two days. Eggs in early stages were in some cases pierced 
through into the yolk with the point of a fine needle at two or three 
points, care being taken not to hurt the germinal disc. These perforated 
eggs were left in 70 per cent, alcohol for one or two days more, and 
were afterwards dehydrated in increasing grades of alcohol. For the 
surface view of the embryo the ventral plate was peeled off the under- 
lying yolk of the preserved egg, and was stained by borax-carmine, 
washed in acidulated alcohol, and imbedded in Canada balsam, after 
dehydration and clarification. 
Eggs which it was proposed to cut into sections were stained with 
borax-carmine or hsematoxylin in toto ; owing to the abundance of the 
yolk the process of section-cutting was very troublesome. Very good 
sections were obtained by the use of the celloidin-paraffin method. 
Investigation of Nectonema4~Mr. H. B. Ward found that the 
resistant cuticle of this worm, which hinders the passage of most fluids, 
and its strong tendency to curl in the killing fluid were great obstacles 
in the successful preservation or sectionizing of specimens. The best 
reagents were found to be a saturated aqueous solution of corrosive 
sublimate and Perenyi’s fluid heated to about 60°. Picro-nitric acid 
gave nearly as good results. The curling of the specimens may be 
largely prevented by straightening the worm gently with the fingers, 
* Quart. Journ. Micr. Sci., xxxiii. (1892) p. 544. 
f Journal Coll. Sci. Imp. Univ. Japan, v. (1892) pp. 56 and 7. 
X Bull. Mus. Comp. Zool., xxiii. (1892) pp. 136 and 7. 
