CILIA 



82 



CILIA 



Heidenhain's "Susa mixture" in his 

 study of ciliogenesis. Mihalik, P. von. 

 (Anat. Anz., 1935, 79, 259-268) preferred 

 the "Susa mixture" for the study of 

 intracellular ciliated cysts in the rabbit 

 oviduct. Meves, Fr. (Arch. mikr. 

 Anat., 1897, 50, 110-114) used Hermann's 

 osmic mixture for the study of sperm 

 flagella of the salamander. Kindred, 

 J. E. (J. Morph. and Physiol., 1927, 

 43, 267-297) recommended Meves' fluid 

 as giving the most constant results in 

 his investigation of ciliogenesis in the 

 pharynx of the frog. Flemming's fluid 

 also gave good results but he did not 

 find 2% osmic acid satisfactory. Ap- 

 plication of Da Fano's modification of 

 Cajal's silver method by Cowdry, E. V. 

 (Anat. Rec, 1921, 22, 289-299) revealed 

 flagella present on the thyroid follicles 

 of the dogfish. The blackened flagel- 

 lum showed up nicely against a yellow- 

 ish background. The collars and fla- 

 gella of sponge choanocytes are difficult 

 to preserve for microscopic study. Rob- 

 bertsou, M. and Minchin, E. A. (Quart. 

 J. Micr. Sci., 1910, N.S._55, 611-640) 

 found Hermann's fluid satisfactory and 

 stained with Heidenhain's iron hema- 

 toxylin and light green. 



Cilia and flagella of protozoa can be 

 demonstrated in fresh preparations by 

 first introducing strands of some ma- 

 terial such as cotton fibers or frayed 

 lens paper beneath the cover glass which 

 will limit activity of the organisms. 

 Maier, H. N. (Arch. f. Protist., 1903, 

 2, 73-179) drew 1% osmic acid and sub- 

 sequently 5% soda solution beneath the 

 cover glass to study cilia and this 

 method is still used. Noland, L. E. 

 (Science, 1928, 67, 535) used a gentian 

 violet stain (20 mg. gentian violet to 

 1 cc. aq. dest ; plus 80 cc. sat. aq. phenol ; 

 20 cc. of 40% formaldehyde; 4 cc. gly- 

 cerin) which he mixed with a drop of 

 ciliates or flagellates in culture. Don- 

 aldson, R. (Lancet, 1917, 2, 571-573) 

 demonstrated protozoa in feces by using 

 0.1 to 0.5% aq. of eosin, or with iodine 

 solutions of various strengths, or with 

 a mixture of the eosin and dilute iodine. 

 These solutions are also good for dem- 

 onstrating cilia and flagella. 



Fixation of flagella by cupro-picro- 

 formol acetic mixture and staining by 

 eosin and phosphomolybdic acid ac- 

 cording to A. C. Hollande's (Arch, de 

 Zool. exper. et gen., 1920, 59, Notes et 

 Revue, 75-77) method is satisfactory. 

 This technique is described in detail by 

 Wenrich, D. H. (McClung's Micr. Tech- 

 nique, 1937, Hoeber, p. 547). 



Relief staining demonstrates ciliary 

 rows and other surface markings on 

 protozoa: B. M. Klien's silver method 



(Zool. Anz., 1926, 67, 160-162) opal blue, 

 china blue, and other stains dried on 

 the protozoa (Bresslau, E., Arch. f. 

 Protist., 1921, 43, 467-480) and uigrosin 

 (Coles, A. C, Watson's Micr. Rec, 

 1927, 23-25). 



Owen, H. M. (Trans. Amer. Micr. 

 Soc, 1947, 66, 50-58) maintains that the 

 "brush" or "flimmer" effect produced 

 on flagella of bacteria by Loeffler's 

 stain is an artifact due to interaction of 

 heavy metals and mordant. He recom- 

 mends a fixative which does not produce 

 the "brush" on flagella (10 parts, 2% 

 osmic acid to 1 part 20% formalin). 



Ciliogenesis can be observed occur- 

 ring in a number of protozoa. Lucas, 

 M. S. (Arch. f. Protist., 1932, 77, 407- 

 423) used vom Roth's platinic chloride 

 fixative followed by Heidenhain's iron 

 hematoxylin stain on smears of Cya- 

 thodinium from the guinea pig intestine, 

 demonstrated formation of new basal 

 bodies and cilia within cysts which later 

 evert upon the protozoan's surface. 



When cilia or flagella are to be studied 

 under the electron microscope, it is 

 necessary, of course, that they be dried 

 under high vacuum. This may intro- 

 duce distortion. A critique on the ad- 

 vantages, limitations, and possible arti- 

 facts of electron microscope techniques 

 has been given by Williams, R. C. 

 (Growth Symposium, 1947, 11, 205-222). 

 Baylor, M. R. B., Nalbandov, A., and 

 Clark, G. G. (Proc. Soc. E.xp. Biol, and 

 Med., 1943, 54, 229-232) studied the 

 sperm head and axial filament. They 

 observed that fresh sperm, dried and 

 dehydrated showed a fraying out of the 

 axial filament into its component fibrils 

 whereas stained sperm did not and they 

 conclude that the stain is sufficiently 

 adhesive to prevent the separation. 

 Brown, H. P. (Ohio J. Sci., 1945, 45, 

 247-301), in his study of flagella, gives 

 his technique for preparation of his 

 material in great detail, including the 

 results from his tests on numerous fixa- 

 tives and stains suitable for use with the 

 electron microscope. Jakus, M. A. and 

 Hall, C. E. (Biol. Bull., 1946, 91, 141- 

 144) studied the trichocyst and cilia of 

 Paramecium and used the shadow-cast- 

 ing technique. 



Microdissection has been used oc- 

 casionally to study the mechanisms of 

 ciliary movement and coordination. 

 The techniques of microdissection are 

 well known from various books and 

 articles. It was used by v. R6nyi, G. 

 (Zeit. f. Anat. u. Entwick, 1926, 81, 

 692-709) to determine if cilia, separated 

 from their basal bodies, were contrac- 

 tile. Carter, G. S. (Proc. Roy. Soc. 

 B, 96, 115-122) made use of this tech- 



