1. Canine Vaginal Cytology
What causes the change in cytology?
How do you take a sample?
What do the cells look like?

2. Canine Vaginal Cytology
This presentation assumes you know the normal canine estrous cycle, the hormones of the estrous cycle and basic canine anatomy.
When you ‘click’, wait a second to see what happens.

3. Typical Call
The cells look kind of round, but with a little angle and the nucleus stains dark.
?????
Dr. Bruce Eilts

4. Typical Result
Just send me the slides.
Dr. Bruce Eilts

5. Demystifying Vaginal Cytology

6. Canine Vaginal Cytology
How do you take a sample?
What causes the change in cytology?
What do the cells look like?

7.                                                                                                                                                                                                                   
                                           
                                         
               
Vaginal Cytology
2. Lift the dorsal aspect of the vulva and insert the swab at the dorsal aspect. Point posterior somewhat and make a ‘dimple’ between the rectum and dorsal vulva.
1. Moisten a long cotton-tipped wooden applicator with a few drops of sterile saline.

8.                                                                                                                                                                                                                   
                                           
                                         
               
Vaginal Cytology
3. Push and rotate the swab in dorsally. It takes more pressure than many realize. If it does not go in easily, back out a little and spin it, as it may be caught on a vaginal fold. Do not go too far ventral or you will go into the bladder.
Good!

9.                                                                                                                                                                                                                   
                                           
                                         
               
Vaginal Cytology
Bad!

10.                                                                                                                                                                                                                   
                                           
                                         
               
Vaginal Cytology
Cells taken from the vestibule or caudal vagina may not represent the true cellular population.
Vestibule

11.                                                                                                                                                                                                                   
                                           
                                         
               
Slide Prep and Stain
Roll the swab out onto a slide. Roll in different ‘streaks’ down the slide.
Label the slide in pencil with the name and date.
Use Dif-Quik stain.
Rinse or put under tap.
10 dips
15 dips
20 dips

12. Canine Vaginal Cytology
How do you take a sample?
What causes the change in cytology?
What do the cells look like?

13. At the end of anestrus and during proestrus, the estrogen from the follicles is rising and peaks just before the beginning of estrus.

14. It is the ESTROGEN that causes the vaginal cytology to change
It is the ESTROGEN that causes the vaginal cytology to change. If there is no estrogen, the vaginal cytology does not change.

15. Without ESTROGEN, (during anestrus and diestrus) the vaginal cytology is non-cornified.

16. Without ESTROGEN, the vaginal epithelium is very thin and the cytology is non-cornified.
With ESTROGEN, the vaginal epithelium is very thick and hyperplastic and the cytology is cornified.
Without ESTROGEN, (even if progesterone is high as in diestrus) the vaginal epithelium is very thin and the cytology is non-cornified.

17. Canine Vaginal Cytology
How do you take a sample?
What causes the change in cytology?
What do the cells look like?

18. Cell types: Cornified vs Non-cornified

19. Anuclear Superficial Intermediate Parabasal
Anuclear cells have an angular cytoplasm and no visible nucleus
Superficial cells have an angular cytoplasm and a non-stippled or pyknotic nucleus
Superficial
Intermediate cells have more angular cytoplasm and a stippled nucleus
Intermediate
Parabasal cells have round cytoplasm and a stippled nucleus
Parabasal

20. Anuclear Superficial Intermediate Parabasal Cornified Non-cornified
Anuclear cells have an angular cytoplasm and a no visible nucleus
Cornified
Superficial cells have an angular cytoplasm and a non-stippled or pyknotic nucleus
Superficial
Intermediate cells have more angular cytoplasm and a stippled nucleus
Intermediate
Non-cornified
Parabasal cells have round cytoplasm and a stippled nucleus
Parabasal

21. Anuclear Superficial Intermediate Parabasal Cornified Non-cornified
This should help you remember the cell names.
Superficial
Intermediate
Non-cornified
Parabasal

22. Anuclear Superficial Intermediate Parabasal Cornified Non-cornified
This should also help you remember what the cells look like.
Anuclear
Cornified
Superficial
Remember, if the nucleus is there, but not stippled, the cell is cornified.
Intermediate
If the nucleus is stippled, the cell is non-cornified.
Non-cornified
Parabasal

23. * * * * S S S A A I * A P I A A * * I * * * * * * * *
Can you find the P’s?
Identify and click.
Can you find the RBC’s?
Identify and click.
Can you find the A’s?
Identify and click.
Can you find the I’s?
Identify and click.
Can you find the S’s?
Identify and click. * * * S S S A A I * A P I A A * * I * * * * * * * *
The resolution of photos makes it difficult to identify the stippling. It is easier looking through the microscope. There is some judgment involved too. Cornified and non-cornified are the big differences to note.

24. During anestrus and diestrus, the cornification is minimal.

25. Vaginal cytology is an estrogen assay
Vaginal cytology is an estrogen assay. If estrogen is low, the epithelium is thin and very few cells, non-cornified cells can be obtained.

26. This animation shows the swab being pushed back and forth, in reality it is rolled. The top is a cartoon and the bottom is a photomicrograph of a non-estrual bitch.
Note the epithelium is thin and not many cells are obtained when they female is non-estrual (anestrus or diestrus).

27. This animation shows the swab being pushed back and forth, in reality it is rolled onto the slide.

28. Notice how few cells are on the slide.

29. Here is glass slide from a non-estrual bitch
Here is glass slide from a non-estrual bitch. Notice that there are not many cells.
Here is photomicrograph from a non-estrual bitch. Notice that there are not many cells, they are non-cornified (see the stippling?), and there is lot of ‘junk’ in the background.

30. During proestrus the cornification rises about 10%/day, until estrus when it reaches 100%. It is 100% cornified throughout estrus. The cornification abruptly drops to <50% on the first day of diestrus.

31. Vaginal cytology is an estrogen assay
Vaginal cytology is an estrogen assay. If estrogen is high, the epithelium is thick and many cornified cells can be obtained.

32. This animation shows the swab being pushed back and forth, in reality it is rolled. The top is a cartoon and the bottom is a photomicrograph of a non-estrual bitch.
Note the epithelium is thick and hyperplastic. Many cells are obtained when the female is estrual (proestrus or estrus).

33. Notice how many cells will be on the slide.

34. Here is a glass slide from an estrual bitch
Here is a glass slide from an estrual bitch. Notice that there are many cells.
Here is a photomicrograph from an estrual bitch. Notice that there are many cells, they are cornified, and the background is clear.

35. Flor ICBAS Lopes & Rocha Bloodhound 2006
‘Flor’ is a bloodhound. She was seen at the University of Porto by Drs. António Rocha and Graça Lopes. Vaginal cytology and progesterone samples were taken. Follow the slides, along with the progesterone values. If you are astute, you see the picture is a male!
Flor
ICBAS
Lopes & Rocha
Bloodhound
2006
Skip this portion and go directly to a self assessment by clicking here.

36. 16/11
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The date is on the top (European style with the day/month). The progesterone (ng/mL) is on the bottom.
A picture of the glass slide lets you see how cellular the sample looks.
Flor
The objective magnification is in the lower left.
20x

37. Follow the slides and see when you think she is estrus and when she should be bred. See when you think she goes out of estrus and into diestrus.

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Flor
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76. Pregnant
Bred on the days listed in red
Whelped 11 pups

77. You can bail out now or go on to a short self assessment.
Hit escape to exit
Left click to continue.

78. What kind of cell is this?
Move you cursor over your answer and left click. You may select them all eventually.
Parabasal
Intermediate
Superficial
Anuclear
Incorrect. Do you see a nucleus? What is the shape of the cytoplasm?
Incorrect. Do you see a nucleus? What is the shape of the cytoplasm?
Incorrect. Do you see a pyknotic nucleus?
Correct. Notice the lack of nucleus and the angular cytoplasm.

79. What kind of cell is this?
Move you cursor over your answer and left click. You may select them all eventually.
Parabasal
Intermediate
Superficial
Anuclear
Incorrect. Do you see the pyknotic nucleus? The cytoplasm is angular.
Incorrect. Do you see the pyknotic nucleus? What is the shape of the cytoplasm?
Correct. You see a pyknotic nucleus and angular cytoplasm.
Incorrect. Notice the pyknotic nucleus.The cytoplasm is angular however.

80. What kind of cell is this?
Move you cursor over your answer and left click. You may select them all eventually.
Parabasal
Intermediate
Superficial
Anuclear
Correct. Notice the stippled nucleus and round cytoplasm.
Incorrect. The nucleus is stippled, but the cytoplasm is not angular.
Incorrect. Do you see the stippled nucleus? It is not pyknotic. The cytoplasm is rounded and not angular.
Incorrect. Do you see the stippled nucleus? It is present. The cytoplasm is rounded and not angular.

81. What kinds of cells are these?
Move you cursor over your answer and left click. You may select them all eventually.
Parabasal
Intermediate
Superficial
Anuclear
Incorrect. Notice the stippled nucleus but the cytoplasm is more angular. A judgment call.
Correct. The nucleus is stippled, and the cytoplasm is angular.
Incorrect. Do you see the stippled nucleus? It is not pyknotic. The cytoplasm is somewhat angular.
Incorrect. Do you see the stippled nucleus? The cytoplasm is Somewhat angular.

82. Are these cells cornified or non-cornified?
Move you cursor over your answer and left click. You may select them all eventually.
Cornified
Noncornified
Incorrect. Notice the stippled nucleus. Cornified cells do not have a stippled nucleus
Correct. The nucleus is stippled.

83. Are these cells cornified or non-cornified?
Move you cursor over your answer and left click. You may select them all eventually.
Cornified
Noncornified
Incorrect. Notice the stippled nucleus. Cornified cells do not have a stippled nucleus
Correct. The nucleus is stippled.

84. Is this cell cornified or non-cornified?
Move you cursor over your answer and left click. You may select them all eventually.
Cornified
Noncornified
Correct. Notice the lack of a nucleus. Non-cornified cells have a stippled nucleus.
Incorrect. Notice the lack of a nucleus. Non-cornified cells have a stippled nucleus.

85. Is this cell cornified or non-cornified?
Move you cursor over your answer and left click. You may select them all eventually.
Cornified
Noncornified
Correct. Notice the pyknotic nucleus. Non-cornified cells have a stippled nucleus.
Incorrect. Notice the pyknotic nucleus. Non-cornified cells have a stippled nucleus.

86. If you saw this slide, what stage of estrus would you place her?
Move you cursor over your answer and left click. You may select them all eventually.
Anestrus
Proestrus
Estrus
Diestrus
Incorrect. Notice all the cells are cornified. The nuclei are pyknotic.
Incorrect. Notice all the cells are cornified. The nuclei are pyknotic. In proestrus there is a mixture of cornified and non-cornified cells.
Correct. Notice all the cells are cornified. The nuclei are pyknotic.
Incorrect. Notice all the cells are cornified. The nuclei are pyknotic. In diestrus there is a mixture of cornified and non-cornified cells.

87. If you saw this slide, what stage of estrus would you place her?
Move you cursor over your answer and left click. You may select them all eventually.
Estrus
Anestrus
Incorrect. Notice some of the cells are non-cornified.
Incorrect. Notice some of the cells are cornified. There are many cells present.
Diestrus
Maybe. Notice about half the cells are cornified. It could be diestrus, but what else could it be?
Proestrus
Maybe. Notice about half the cells are cornified. It could be proestrus, but what else could it be?

88. If you saw this slide, how would you tell if she is proestrus or estrus?
Run Estrogen Assay
Move you cursor over your answer and left click. You may select them all eventually.
No way to tell
Incorrect. Estrogen assays are difficult and not rewarding for this.
Incorrect. Remember your physiology. You can tell.
Run Progesterone Assay
Take another swab in 2 days.
Correct. Progesterone will be low in proestrus and high in diestrus. This will tell us today. Any cheaper way to tell?
Correct. If she is in proestrus, it will become more cornified. If she is in diestrus it will become less cornified. Any more precise way to tell?

89. Session Complete. Thanks for viewing this presetation
Session Complete. Thanks for viewing this presetation.I hope it helped clarify canine vagnal cytology.