Isolation of salivary glands from ticks Ixodes ricinus

   The procedure is based on an  article by L. Paulin Maldonado-Ruiz and  his
   team (1996). The salivary glands were prepared from 14 females into 2 test
   tubes. The  selected  individuals were  females,  due to  better  handling
   compared to small nymphs.

   Reagents

   • 0.5% NaClO

   • 70% Ethanol

   • PBS (phosphate buffer) pH 7.4

   • dH2O

   Instruments and laboratory equipment

   • Binocular microscope (brand)

   • Scalpel, tweezers, needle

   • Common laboratory equipment:  Eppendorf test tubes, automatic  pipettes,
   tips, petri dishes, parafilm,

   Method

   Subjects were  placed one  at a  time on  a petri  dish, where  they  were
   sterilized with  0.5% sodium  hypochlorite.  Subsequently, wash  with  70%
   ethanol and wash with dH2O before the preparation itself. The subject  was
   placed on a piece of parafilm, which was spread on the surface of a  petri
   dish, and then the tick was  dripped with PBS buffer. The preparation  was
   performed under a binocular  microscope, where it  was performed from  the
   front part,  in order  to avoid  as much  as possible  contamination  with
   intestinal components.  The  dorsal  integument was  first  dissected  and
   removed, and  then  the  salivary  glands  (anatomy  see  Figure  5)  were
   transferred to a tube  with PBS buffer. Thus,  the samples were stored  at
   -20 ° C.

   Location of salivary glands in the tick Ixodes ricinus (URL5), modified