2088 Journal of Applied Microscopy 



The Technique of Biological Projection and Anesthesia of 



Animals. 



Copyrighted. 



IX. THE ANESTHESIA OF ANIMALS.— Continued. 



To produce a state of hypnosis, or complete anesthesia, in animals by the 

 use of chloretone, either one of four methods of administration may be used, or 

 two of them may be combined, as each case may demand. 



1. Place the animals in water sufficient to cover them and add chloretone 

 stock solution, i. e., one per cent, solution in tap water, slowly drop by drop, or 

 in larger quantities, until the desired state is attained. 



2. Place the animals in a dilute solution of definite strength, or in the full 

 strength stock solution. Use tap water, or water in which the animals naturally 

 live, for making dilute solutions from the stock solution. 



3. Inject stock solution into the animal's stomach by means of a pipette or 

 syringe. 



4. Feed the animal with a proper dose of the chloretone crystals in capsules 

 or sugar-coated tablets. 



Protozoa : Amoeba. — Although these animals do not usually move with 

 such rapidity as to require anesthesia for their careful study in the live state, 

 the interesting phenomena which they present under the influence of chloretone 

 justify a description of the method. 



Amoebae are easily cultivated in the laboratory in shallow, flat-bottomed, 

 loosely covered vessels, such as large crystallizing or culture dishes. If the 

 amoeba stock is obtained from the surface of the mud in a pond or ditch, care 

 should be taken not to transfer enough mud to completely cover the bottom of 

 the dish. Areas free from the coarser sediment should appear after it has 

 settled. Let such a culture stand quietly for a few days, not in direct sunlight 

 and free from snails, which probably destroy the amoebae. When ready for 

 study, take a clean watch-glass and a clean rubber bulb pipette having a good 

 point of medium or small bore, and scratch the clear areas vigorously with the 

 point of the pipette and at the same time allow the pipette to fill with water con- 

 taining fine mud, microscopic plants, and amoebae. Transfer this water and its 

 floating contents at once to the watch-glass. The amoeba; adhere to the surface 

 of the glass, therefore vigorous scratching is necessary to loosen them, and they 

 quickly settle upon and adhere to the inside of the pipette, if not transferred 

 immediately to the watch-glass. 



Place the watch-glass under a one-half or three-quarter inch objective, and 

 focus carefully on the surface of the glass at the bottom of the water. In a few 

 minutes the amoebae will be found adherent to and flowing over the surface of 

 the glass, provided that the glass and water are at ordinary room temperature, 

 or a few degrees above. To get rid of the dirt, other organisms, and nearly all 

 of the water, pour it off by a quick tilting of the watch-glass and the amoebae 

 will be found, on again examining with the microscope, to be moving about in 



