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Then, while a flngor is held against the end, the plunger is pushed out 
again until the air around it has been entirely replaced by water* A 
5«4im* column of water is left at the end of the plunger* 
The plastic syringe can be prepared in much the same way as the 
glass syringe when a lubricant seems necessary* Ordinarily the water 
will be present after the rinsing operation described later. The plunger 
should be pushed out as far as it will go* 
Tap water has been found satisfactory, but in some localities distilled 
water may be better* Physiological salt solution must not be used with the 
glass syringe, because it cannot be thoroughly rinsed and, when the water 
dries up, the salt residue freezes the plunger in the tip and corrodes 
the syringe parts* 
Preparing the Queen 
The queen is made to walk into a tube similar in site and construc- 
tion to the queen holder* When she reaches the partly closed end she 
begins backing up, and if the queen holder is quickly put in place she 
usuedly backs into it* The stopper is pushed in after her until her 
abdomen protriides from the sme^ll end of the holder and she cannot move 
about readily* For best results only about the last three segments of 
the abdomen should protrude* 
The three-'way stopcock is turned so that carbon dioxide will flow 
throu^ the queen holder* The queen breathes heavily for a few minutes 
and then gradually becomes quiet* If her abdomen expands abnormally, 
the gas is being forced into her abdominal air sacs, an indication that 
the flow is too strong* Unless very severe this action seems to do no 
harm, although it makes insemination difficult* The holder is withdrawn 
into its mounting block by pulling on the carbon dioxide supply tube, 
and tightened in place by the set screw* The dorsal part of the queen 
should be at the right of the operator* The queen holder should be about 
30° from vertical, with the upper end leaning to the right (fig* 4)* 
The queen is usually quiet by the time she is completely mounted, 
and the holding hooks can be put in place. This is done under low-power 
magnification* First one hook and then the other is inserted into the 
sting chamber, and the abdominal plates sure pulled apart* With the left 
hand the sting depressor is used to hold the sting down while the sting 
hook is placed in the triangular area between the bases of the sting 
lancets and the sting hook is left in this position to prevent umiacessary 
drying of the delicate tissues while the syringe is being loaded. 
Filling the Syringe 
The microscope is withdrawn slightly and the syringe placed in its 
box, with care not to break the tip* This is done with greatest safety 
by depressing the spring with forceps while the syringe is being Inserted* 
The microscope and syringe are then so adjusted that the end of the syringe 
is in focus* 
