canine vaginal cytlogy

Canine Vaginal Cytology

• What causes the change in cytology?

• How do you take a sample?• What do the cells look like?


• 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.

Typical CallThe cells look

kind of round, but with a little angle and the nucleus stains dark.

Dr. Bruce Eilts

?????

Typical Result

Dr. Bruce Eilts

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?

Vaginal Cytology

1. Moisten a long cotton-tipped wooden applicator with a few drops of sterile saline.

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.

http://www.vetmed.lsu.edu/eilts/Multimediafiles/Sound/Video/Canine%20Cytology.mpg

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.

Vestibule


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 dips15 dips 20 dips

Rinse or put under tap.


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 non-cornified.

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.

Cell types:Cornified vs

Non-cornified

Parabasal

Intermediate

Superficial

Anuclear

Parabasal cells have round cytoplasm and a stippled nucleus

Intermediate cells have more angular cytoplasm and a stippled nucleus

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

Anuclear cells have an angular cytoplasm and no visible nucleus

Parabasal

Intermediate

Superficial

Anuclear

Cor

nifie

dN

on-c

orni

fied

Parabasal cells have round cytoplasm and a stippled nucleus

Intermediate cells have more angular cytoplasm and a stippled nucleus

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

Anuclear cells have an angular cytoplasm and a no visible nucleus

Parabasal

Intermediate

Superficial

Anuclear

Cor

nifie

dN

on-c

orni

fied

This should help you remember the cell names.

Parabasal

Intermediate

Superficial

Anuclear

Cor

nifie

dN

on-c

orni

fied

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

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

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

P

I

SA

SA

AA

I

*

*

*

*

*

*** *

*

*

*

I

S

A

*

**

Can you find the P’s?Identify and click.Can you find the I’s?Identify and click.Can you find the S’s?Identify and click.

Can you find the A’s?Identify and click.Can you find the RBC’s?Identify and click.

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.

http://www.vetmed.lsu.edu/eiltslotus/theriogenology-5361/vaginal_cells.htm

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 non-cornified (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.

http://www.vetmed.lsu.edu/eiltslotus/theriogenology-5361/vaginal_cells.htm

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).

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. 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!

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

16/11

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Flor

20x

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.

The objective magnification is in the lower left.

• 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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.15 - - - .78 .87 3.2 8.2 - 34 - - -

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Flor

100x

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 Intermediate Superficial Anuclear

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.

Incorrect. Do you see a nucleus? What is the shape of the cytoplasm?

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



Incorrect. Do you see the pyknotic nucleus? The cytoplasm is angular.

Correct. You see a pyknotic nucleus and angular cytoplasm.

Incorrect. Notice the pyknotic nucleus.The cytoplasm is angular however.

Incorrect. Do you see the pyknotic nucleus? What is the shape of the cytoplasm?




Correct. Notice the stippled nucleus and round cytoplasm.

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.

Incorrect. The nucleus is stippled, but the cytoplasm is not angular.


What kinds of cells are these?


Incorrect. Notice the stippled nucleus but the cytoplasm is more angular. A judgment call.

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.

Correct. The nucleus is stippled, and the cytoplasm is angular.


Are these cells cornified or non-

cornified?

Cornified Noncornified

Incorrect. Notice the stippled nucleus. Cornified cells do not have a stippled nucleus

Correct. The nucleus is stippled.


Is this cell cornified or non-cornified?


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.


Is this cell cornified or non-cornified?


Correct. Notice the pyknotic nucleus. Non-cornified cells have a stippled nucleus.

Incorrect. Notice the pyknotic nucleus. Non-cornified cells have a stippled nucleus.


If you saw this slide, what stage of estrus would you

place her?

Anestrus Proestrus Estrus Diestrus

Incorrect. Notice all the cells are cornified. The nuclei are pyknotic.

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.

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


If you saw this slide, what stage of estrus would you

place her?Anestrus

Proestrus

Estrus

Diestrus

Incorrect. Notice some of the cells are cornified. There are many cells present.

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

Maybe. Notice about half the cells are cornified. It could be diestrus, but what else could it be?

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?

No way to tell

Take another swab in 2 days.

Run Estrogen Assay

Run Progesterone Assay

Incorrect. Remember your physiology. You can tell.

Incorrect. Estrogen assays are difficult and not rewarding for this.

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?


Session Complete.Thanks for

viewing this presetation.I

hope it helped clarify canine

vagnal cytology.

canine vaginal cytlogy

Documents

pyknotic nucleusintermediate

superficial

canine vaginal

vaginal cytology

superficial

dorsal aspect

angular cytoplasm

animation