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The research on ciliates, flagelates and opalinates have been widespread by the …


Biology Articles » Protistology » Improvement of silver impregnation technique (protargol) to obtain morphological features of protists ciliates, flagellates and opalinates » Methods

Methods
- Improvement of silver impregnation technique (protargol) to obtain morphological features of protists ciliates, flagellates and opalinates

The ciliates Tracheloraphis phenicoptera, Coleps sp. and Diophrys appendiculata were collected from Baía de Guanabara, Rio de Janeiro; Lacrymaria olor and Spirostomum minus were collected in bromeliad; Colpoda cucullus was collected, in form of cysts, from the filter in the air conditioner of the laboratory. Paramecium aurelia and Euplotes woodruffi were collected from Lagoa de Piratininga in Niterói. Fabrea salina was collected from Lagoa de Araruama in Rio de Janeiro. Trichonympha sp. was isolated from digestive apparatus of termites. The opalinates were isolated from the digestive apparatus of tadpoles Physalaemus signifer.

The ciliates were cultivated in petri dishes with rice or wheat grains, except the opalinates and flagellates. The fixatives used were: 2% Osmiun tetroxide (T. phenicoptera), alcoholic-Bouin (D. appendiculata and Coleps sp.), Stieves' solution(L. olor, S. minus, C. cucullus, P. aurelia, E. woodruffi and F. salina) and 2,5% glutaraldehyde (Trichonynpha sp. and the Opalinata).

Alcoholic-Bouin: 150 ml 80% alcohol + 60 ml formaldehyde PA + 1 g of picric acid + 15 ml glacial acetic acid. This is an excellent fixative, one of its advantage is facilitating attaching the ciliates in the bottom of the clock glass, facilitating the subsequent washes.

Solution of Stieves: (modified by Foissner, 1991); 3,8 ml of aqueous mercury chloride (dissolve 30 g HgCl2 in 500 ml of hot distilled water) + 2 ml of commercial formaldehyde PA + 0,3 ml of concentrated glacial acetic acid. freshly prepared; the components can be stored.

Glycerinated-albumin of Mayer: For the preparation of the Glycerinated-Albumin of Mayer is recommended the use of two chicken eggs. Remove the albumin of the egg and filter through a nylon filter and mix 30 ml of the filtered albumin with 20 ml of glycerin; add 3 drops of formaldehyde PA to avoid the contamination of the material.

1% hydroquinone: place 2,5 g of sodium sulfite in 50 ml of distilled water, dissolve, and later add 0,5 g of hidroquinone.


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