Demonstration of intranuclear vesicles in blast cells in lymph nodes with different fixatives and staining techniques

Abstract

For the demonstration or intranuclear vesicles, iodine vapor, formalin vapor, and methyl alcohol fixations give equivalent results when followed by staining with the May-Grünwald Giemsa technique. Neutral buffered formalin and May-Grünwald Giemsa staining demonstrate fewer intranuclear vesicles than the first-mentioned combinations. Iodine vapor or formalin vapor fixations followed by Mayer's haematoxylin staining show greater numbers of nuclei with vesicles.Formalin fixes nuclei slowly, which allows nuclear contraction and thus the squeezing-out of vesicles. Haematoxylin stains protein, while May-Grünwald Giemsa technique does not stain proteins but deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). It therefore follows that the demonstration of vesicles in blast cell nuclei depends on the fixatives and staining methods used.

Protoplasmic fragments in haematopoietic tissues and analysis of intranuclear vesicles in lymph node blast cells of the rabbit

“…The free vesicles were usually round to oval in shape (Figs. 4,5,13,17,20) although sometimes elongated ( Fig. 9, varied in size (Fig.…”

“…the long axis was no more than 3 X the short axis, were analyzed since they make up about 77% of all blast cell nuclei (14). Intranuclear vesicles were defined as spherical pale or non-staining areas, 1 p or greater in diameter, and enclosed for more than SOYo of their circumference (17).…”

“…10-12, [14][15][16][17]19). Most commonly, these cells were lymphocytes of different sizes and blast cells.…”

“…The large numbers of other cellular particles suggest that they too should be examined. Free protoplasmic fragments have been observed in haematopoietic tissues prepared by a variety of techniques, including impression methods (23, 23, smears (19, 23), needle aspirations (17,23), light microscopic preparations (19, 23, 23, electron microscopic preparations (3, 23), and cultures of living cells (3; unpublished observations in this laboratory). We are confident that the observations reported in this paper cannot be artefacts.…”

“…The present paper is concerned with (1) the morphology of certain free protoplasmic fragments and examination of haemopoietic cells in an attempt to understand the origin of the fragments and (2) an analysis of the size and frequency of occurrence of intranuclear vesicles (17) in blast cells devoid of cytoplasm in the mesenteric lymph nodes of rabbits.…”