CILIA 



81 



CILIA 



E. L. (Quart. J. Micr. Sci., 1942, 83, 

 245-258) as a new technique with slight 

 modifications. Lucas, A. M. (J. 

 Morph., 1931, 51, 147-193) used sodium 

 borate to which was added a trace of 

 iodine or eosin. The cells swelled as 

 they separated. Poska-Teiss, L. (Zeit. 

 f. wiss. Mikr., 1934, 51, 238-243) used 

 Ranvier's 1/3 alcohol method to isolate 

 the cells. Loginoff, W. J. (Anat. Anz., 

 1911, 38, 353-361) who worked with 

 horses, cows and sheep, discovered that 

 preliminary fixation of ciliated cells of 

 the trachea for 30 min. in 1% formalin 

 followed by 1/3 alcohol for 24 hrs. gave 

 well preserved cells which could be 

 stained under the coverglass with picro- 

 carmiue. 



In vitro explants of ciliated epithelium 

 show, eventually, dedifferentiation and 

 loss of cilia. More often short term 

 studies on surviving cells have been 

 resorted to. Isolated cells from the 

 frog's palate under tissue culture condi- 

 tions beat continuously since they are 

 no longer regulated by the nervous sys- 

 tem and are suitable for testing pur- 

 poses (Ishikawa, S., ActaDermat., 1927, 

 9, 339-364). Verne, J. (Compt. Rendu 

 Assoc. Anat., 1932, 27, 564-578) using well 

 known tissue culture techniques cul- 

 tivated lung tissue of 15 to 17 day old 

 chick embryos. In this case the cil- 

 iated epithelium did not undergo meta- 

 plasia but remained active. Proetz, A. 

 W., and Pfingsten, M. (Arch. Otolar- 

 yng., 1939, 29, 252-262) combined cul- 

 ture of guinea-pig nasal epithelium with 

 motion picture photography. 



Umeda, T. (Acta Dermat., 1930, 1, 

 13-38) tested some 700 dyes for their 

 vital staining characteristics and found 

 that thionin blue and Nile blue gave 

 satisfactory results with frog ciliated 

 epithelium. Vital methylene blue has 

 frequently been used to stain lining 

 ciliated cells. Carter, G. S. (Brit. J. 

 Exp. Biol., 1926, 4, 1-26) observed that 

 it was the region of the ciliary rootlets 

 which showed granules staining strongly 

 with this dj'e and Coonfield, B. R. 

 (Biol. Bull., 1936, 70, 400-471) found 

 the greatest staining in Nephthys where 

 the rootlets converged near the nucleus. 



Most of the usual histologic or cyto- 

 logic techniques will demonstrate cilia. 

 Cilia are stained chiefly with acid dyes 

 but may hold Heidenhain's iron hema- 

 toxylin if the tissues have been only 

 lightly destained with iron alum. 

 When staining the very short sensory 

 ciliary hairs on olfactory epithelium, 

 the iron hematoxylin without counter 

 stain or only a light counter stain makes 

 them stand out quite well. 



A cuticular border is present on some 



types of ciliary cells and absent on 

 others. It is refractile to most stains 

 and oftentimes appears as a very nar- 

 row clear space between the line mark- 

 ing the surface of the cuticle and the 

 underlying cell membrane. If the over- 

 lying mucus is stained with mucicar- 

 mine, the outer surface of the cuticle 

 is clearly delimited. Frequently, the 

 lower margin of a nonciliated striated 

 cuticular border looks like a row of basal 

 bodies and Heidenhain's iron hema- 

 toxj'lin gives the best differentiation. 

 Likewise it is the stain usually chosen 

 to show flagellate diplosomes and stages 

 in ciliogenesis. 



Ciliary rootlets are as difficult to 

 demonstrate in the stained as in the 

 living condition. In order to see them 

 clearly it is important to select the right 

 kind of cell — one which has a clear 

 protoplasm containing a minimum of 

 pigmented opaque or refractile gran- 

 ules. Even living unstained ciliated 

 cells of the mammalian nose when iso- 

 lated on a slide show clearly the root- 

 lets in the narrow clear zone of proto- 

 plasm just beneath the cell membrane 

 and in relatively transparent cells they 

 can be observed converging toward the 

 nucleus as an inverted truncated cone. 

 In the latero-frontal ciliated cells of 

 the mollusc gill the cilia form two rows 

 transversely across the surface of the 

 cells so that the rootlets lie in rows one 

 behind the other and under these condi- 

 tions they show up very easily. Grave, 

 C, and Schmitt, F. O. (J. Morph. and 

 Physiol., 1925, 40, 479-515) recom- 

 mended the Ehrlich-Biondi triple mix- 

 ture as a counterstain to Heidenhain's 

 iron hematoxylin and this works very 

 well although sometimes more than one 

 attempt is necessary to obtain a satis- 

 factory mixture of Ehrlich-Biondi 

 stain. 



Considerable latitude is permitted in 

 the use of fixatives. Allen's Bj and Bu 

 are good fixatives although the alveolar 

 appearance that picric-containing fixa- 

 tives give to the appearance of the cyto- 

 plasm may be a disturbing factor. Del- 

 linger, O. P. (J. Morph., 1909, 20, 171- 

 209) made an informative comparative 

 study of the fixatives and the basic con- 

 stituents which go into them. He found 

 that HgCli emphasized fibrillar struc- 

 tures and enhanced the staining and so 

 fixatives such as Zenker's, with formalin 

 or acetic acid, are often used for preser- 

 vation of ciliated tissues. These are 

 vigorous fixatives and may result in 

 some cell distortion. Bellinger recom- 

 mended 0.4 to 2% osmic acid as best for 

 cilia. Thanhoffer, L. von. (Zeit. Anat. 

 Entwickl., 1929, 90, 713-724) employed 



