Cover slips acid washed

Should the stain be unsatisfactory, the slide can be destained by placing it in xylene to remove the cover slip or immersion oil and then placing it in 50% alcohol for 10 min to hydrate the slide. Destain the slide in 10% acetic acid in water for several hours, and then wash it thoroughly first in water and then in 50 and 70% alcohols. Place the slide in stain for 8 min, and then complete the stain procedures. It is helpful to eliminate or shorten the destain step. 

A microscopic preparation is made, as described later, of cotton or flax fibres obtained from white fabrics repeatedly washed. These fibres are treated for about 1-2 minutes with the iodine solution, which is then eliminated by careful and complete absorption with thin strips of filter-paper.1 One or two drops of the acid prepared as above are added, the cover-slip placed and the preparation observed under the microscope. If the concentration of the acid is correct, the flax and cotton fibres are coloured blue without being deformed. If, however, the coloration is feeble and tends to red, the acid is too dilute, while if the fibres swell, it is too concentrated. 

Rinse the slides in acid HBSS, and stain for 5 min in 10% Giemsa Wash in pH 6.8 phosphate buffer and mount the slides in the same buffer under a cover slip. Following photography (if required), destain the slides in analar methanol for 5 min, and air-dry. 

Arabidopsis thaliana seedlings were grown using a 1 x 10-4 M Rhodamine-6G aqueous solution on a filter paper of glass fiber. For the preparation of microscopic specimens, Arabidopsis thaliana roots were crushed between two glass slides after soaking in 1M HC1 in a hot-water bath for 10 min, and washed with 45% acetic add and 3 1 methanol/acetic acid. The preparation was sealed in nujol mull with a cover slip. 

Acid washed cover slips Prepare cover slips by boiling in 0.2A HC1 for a few minutes, then rinsing in distilled water. Store the cover slips m 96% ethanol. Before use, dry them with a tissue. The use of the acid-washed cover slip minimizes loss of cells by adhesion to the cover slip. 

Add an acid-washed cover slip and press down over filter paper very gently (it is not necessary to squash). 

As soon as the alcohol has dried, immediately add 9.5 pL of probe cocktail to each slide see Note 5). Add an acid-washed 22 x 22 mm cover slip, trying to avoid air bubbles, and ring with rubber solution. 

Immediately prior to use mix an equal volume of silver amplification solution A with solution B and add 500 pL to the slide. At this point the slide may be covered with a glass cover slip and the silver deposition monitored under the microscope (although light exposure hastens silver deposition). Stop the reaction by washing in water followed by 1% acetic acid for 2 min. 

Linseed Oil.—The best quality of oil is used, and this is purified by digesting it in partially-diluted sulphuric acid for some hours, at a temperaturo of about 212 , allowing the impurities to subside, and then washing away the add with repeated additions of het water. The oil after this treatment is pale and turbid, and, if the edulcoi ation is complete, there is scarcely any odor. By rest the oil clarifies, and has then a pale lemon color. It now dries much more rapidly than before. Tor an experiment, four drops of oil were exposed on a slip of glass the first wae ordinary Baltic linseed oil the second was East India oil the third and fourth were two distinct oils which had been purified by means of sulphuric acid. In the course of a fortnight, the first was colorless and rather viscid, but no skin or pellicle had formed on its surface the second was colorless, but bad no apparent viscidity the third and fourth were not only colorless, but a tenacious film covered the surface of each. 

Spills of ammonia present two hazards. Laboratory workers may slip in the spill or be overcome by the vapors, especially if the ammonia is a concentrated solution. Thus, the worker handling the spill should wear breathing apparatus and other people should remain a safe distance away. The spill is covered with sufficient spill mix to absorb all of the liquid and maintain a dry layer of spill mix on top. The mixture is scooped into a container and transported to the fume hood. In the fume hood it is slowly added to a plastic pail of cold water. The liquid is neutralized by the addition, with stirring, of either solid sodium bisulfate or 5% aqueous hydrochloric acid. The aqueous solution is decanted into the drain and the solid, sand/bentonite, disposed as normal garbage or washed with water, dried, and reused... 

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