| Canine
        Vaginal Cytology 
   p 32
 Technique
 
                Moisten a cotton swab with 1 to 2 drops sterile
            saline. Open the vulvar lips, pull the vulva doraslly, insert the swab dorsally 
				and posterior, then up and over
            the pelvic brim and into anterior vagina. If you do not pass the swab
            far enough, you will get vestibular cells and result in false cornification. If
            you pass the swab too ventral, you may enter the bladder and get a
            falsely non-cornified smear.
                  Roll the swab firmly onto a slide.
                    Stain the slide using DifQuik stain, 10 dips in
            A, 15 dips in B, and 20 dips in C. You may also use new methylene
            blue stain.
                      Read the slide under low power first to
            establish the trend of cellularity and cell types. Move to a higher
            power to establish the cell types. View several fields to get an
            overall visual idea of the percentage of cornified cells. | 
                
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                  |  Click on the movie icon, then right click on the movie or "Open
        It" and "OK" to see
 a Vaginal Cytology Exam performed
 | 
                
                  | Vaginal
        Cytology Cell Types | 
                
                  | Non-cornified 
                      Parabasal cells have a large stippled nucleus
            and a rounded cytoplasm The nucleus is large compared to the
            cytoplasm.  Intermediate cells have a have a stippled
            nucleus and more cytoplasm than parabasal cells. The cytoplasm may
            even become angular.  
 
 | Cornified Superficial cells have a pyknotic nucleus and
            angular cytoplasm.  There is no stipling in the nucleus.
                       
 
 Anuclear cells have no visible nucleus and
            angular cytoplasm.  
 | 
                      
                        | Remember
        'PISA' 
 
  
						
						 
						Click picture above to enlarge the 
						cells. | 
                      
                        | Changes
        during the estrous cycle 
   
                            When no estrogen is present (anestrus and
            diestrus, the vaginal wall is very thin and is comprised of
            noncornified cells. In anestrus there will be very few cells and
            what you see will be debris and non-cornified cells.
                              When estrogen rises during proestrus, the
            vaginal epithelium becomes hyperplastic and more cornified. During
            proestrus the percent of cornified cells increases by about 10%/day
            until you see  about 100% cornification during estrus. You may also see
            RBCs during proestrus. | Thin vaginal epithelium during
        anestrus. 
  | 
                        
                          | Estrus 
                              During estrus the vaginal epithelium is very
            thick. You will see almost 100% cornification. The smear will look
            the same from the first day of estrus of the last day of estrus, You
            cannot tell which day of estrus the bitch is in based on vaginal
            cytology. There may however, be some sheeting of cells during the
            last 1-2 days of estrus. The vaginal wall is so thick during estrus
            that PMNs do not cross the epithelium. This makes the background
            look very 'clean'. | 
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                          | Estrus smear with 100%
        cornification and a clear background.
  | Sheets of cells during the end
        of estrus.  | 
                        
                          | Diestrus 
                              On the first day of diestrus the cells in the
            swab abruptly change to around 50% non-cornified. This day that the
            smear changes from 100% cornified cells to 50% non-cornified cells
            is denoted as the first day of cytologic diestrus. You may see an
            influx of PMNs at this time to help clean up all the cellular
            debris. | Abrupt change
        to non-cornified cells on the first day of diestrus.   |