What causes the change in cytology? How do you take a sample? What do the cells look like?

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.

Canine Vaginal Cytology

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

Typical Result
Just send me the slides.

Demystifying Vaginal Cytology

 How do you take a sample? What causes the change in cytology? What do the cells look like?

Canine Vaginal Cytology

Vaginal Cytology
2. Lift the dorsal aspect of the 1. Moisteninsert the swab at vulva and a long cottontipped wooden applicator with the dorsal aspect. Point a few drops of sterile and posterior somewhat saline. make a dimple between the rectum and dorsal vulva.

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!

Vaginal Cytology

Bad!

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

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.

10 dips

15 dips

20 dips

Rinse or put under tap.

 How do you take a sample? What causes the change in cytology? What do the cells look like?

Canine Vaginal Cytology

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

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

Without ESTROGEN, (during anestrus and diestrus) the vaginal cytology is noncornified.

Without ESTROGEN, the vaginal epithelium is very thin and the cytology is noncornified.

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 noncornified.

 How do you take a sample? What causes the change in cytology? What do the cells look like?

Canine Vaginal Cytology

Cell types: Cornified vs Non-cornified

Anuclear cells have an angular cytoplasm and no visible nucleus

Anuclear
Superficial Intermediate Parabasal

Superficial cells have an angular cytoplasm and a non-stippled or pyknotic nucleus

Intermediate cells have more angular cytoplasm and a stippled nucleus

Parabasal cells have round cytoplasm and a stippled nucleus

Superficial cells have an angular cytoplasm and a non-stippled or pyknotic nucleus

Intermediate cells have more angular cytoplasm and a stippled nucleus

Superficial Intermediate Parabasal

Parabasal cells have round cytoplasm and a stippled nucleus

Non-cornified

Cornified

Anuclear cells have an angular cytoplasm and a no visible nucleus

Anuclear

This should help you remember the cell names.

Superficial Intermediate Parabasal

Non-cornified

Cornified

Anuclear

Superficial
Remember, if the nucleus is there, but not stippled, the cell is cornified.

If the nucleus is stippled, the cell is non-cornified.

Parabasal

Non-cornified

Intermediate

Cornified

This should also help you remember what the cells look like.

Anuclear

* * * *

Can you find the As? Can you find thePs? RBCs? Is? Ss? Identify and click. Identify and click.

A
*

S
I

A
A

P 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.

During anestrus and diestrus, the cornification is minimal.

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

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).

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

Notice how few cells are on the slide.

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 noncornified (see the stippling?), and there is lot of junk in the background.

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.

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

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 nonestrual bitch. Note the epithelium is thick and hyperplastic. Many cells are obtained when the female is estrual (proestrus or estrus).

Notice how many cells will be on the slide.

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.

Flor

ICBAS Lopes & Rocha Bloodhound 2006

Flor is a bloodhound. She was seen at the University of Porto by Drs. Antnio Rocha and Graa 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!

Skip this portion and go directly to a self assessment by clicking here.

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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.

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 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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Pregnant
Bred on the days listed in red Whelped 11 pups

You can bail out now or go on to a short self assessment.

Hit escape to exit Left click to continue.

What kind of cell is this?

Parabasal Incorrect. Do you see a nucleus? What is the shape of the cytoplasm? Intermediate Incorrect. Do you see a nucleus? What is the shape of the cytoplasm? Superficial Incorrect. Do you see a pyknotic nucleus?

Move you cursor over your answer and left click. You may select them all eventually. Anuclear Correct. Notice the lack of nucleus and the angular cytoplasm.

What kind of cell is this?

Parabasal Incorrect. Do you see the pyknotic nucleus? The cytoplasm is angular. Intermediate Incorrect. Do you see the pyknotic nucleus? What is the shape of the cytoplasm? Superficial Correct. You see a pyknotic nucleus and angular cytoplasm.

Move you cursor over your answer and left click. You may select them all eventually. Anuclear Incorrect. Notice the pyknotic nucleus.The cytoplasm is angular however.

What kind of cell is this?

Parabasal Correct. Notice the stippled nucleus and round cytoplasm. Intermediate Incorrect. The nucleus is stippled, but the cytoplasm is not angular. Superficial Incorrect. Do you see the stippled nucleus? It is not pyknotic. The cytoplasm is rounded and not angular.

Move you cursor over your answer and left click. You may select them all eventually. Anuclear Incorrect. Do you see the stippled nucleus? It is present. The cytoplasm is rounded and not angular.

What kinds of cells are these?

Parabasal Incorrect. Notice the stippled nucleus but the cytoplasm is more angular. A judgment call. Intermediate Correct. The nucleus is stippled, and the cytoplasm is angular. Superficial Incorrect. Do you see the stippled nucleus? It is not pyknotic. The cytoplasm is somewhat angular.

Move you cursor over your answer and left click. You may select them all eventually. Anuclear Incorrect. Do you see the stippled nucleus? The cytoplasm is Somewhat angular.

Are these cells cornified or non-cornified?

Move you cursor over your answer and left click. You may select them all eventually. Cornified Incorrect. Notice the stippled nucleus. Cornified cells do not have a stippled nucleus Noncornified Correct. The nucleus is stippled.

Are these cells cornified or non-cornified?

Move you cursor over your answer and left click. You may select them all eventually. Cornified Incorrect. Notice the stippled nucleus. Cornified cells do not have a stippled nucleus Noncornified Correct. The nucleus is stippled.

Is this cell cornified or non-cornified?

Move you cursor over your answer and left click. You may select them all eventually. Cornified Correct. Notice the lack of a nucleus. Noncornified cells have a stippled nucleus. Noncornified Incorrect. Notice the lack of a nucleus. Noncornified cells have a stippled nucleus.

Is this cell cornified or non-cornified?

Move you cursor over your answer and left click. You may select them all eventually. Cornified Correct. Notice the pyknotic nucleus. Noncornified cells have a stippled nucleus. Noncornified Incorrect. Notice the pyknotic nucleus. Noncornified cells have a stippled nucleus.

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 Incorrect. Notice all the cells are cornified. The nuclei are pyknotic. Proestrus Incorrect.
Notice all the cells are cornified. The nuclei are pyknotic. In proestrus there is a mixture of cornified and non-cornified cells.

Estrus Correct. Notice all the cells are cornified. The nuclei are pyknotic.

Diestrus Incorrect. Notice

all the cells are cornified. The nuclei are pyknotic. In diestrus there is a mixture of cornified and non-cornified cells.

If you saw this slide, what stage of estrus would you place her?
Anestrus Incorrect.
Notice some of the cells are cornified. There are many cells present.

Move you cursor over your answer and left click. You may select them all eventually.

Estrus
Incorrect. Notice some of the cells are non-cornified. 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?

If you saw this slide, how would you tell if she is proestrus or estrus?
Move you cursor over your answer and left No way to tell click. You may select them all eventually. Incorrect.
Remember your physiology. You can tell.

Run Estrogen Assay Incorrect.

Estrogen assays are difficult and not rewarding for this.

Take another swab in 2 days. 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?

Run Progesterone Assay Correct.

Progesterone will be low in proestrus and high in diestrus. This will tell us today. Any cheaper way to tell?

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