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Jos - Cytology DIY?

Equine-Reproduction.com Bulletin Board » Breeding Methods » Jos - Cytology DIY? « Previous Next »


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hedgerow pony farm
Breeding Stock
Username: Lotsofponies

Post Number: 214
Registered: 03-2009
Posted on Monday, August 22, 2011 - 10:59 am:   Edit Post Delete Post    Move Post (Moderator/Admin Only)

It seems like vets are not to eager to do cytology. Last month a friend bred his mare AI, turn it all over to the vet, the mare was at the clinic for a week. From the beginning he asked her to do a cytology, didn't do it because no evidence of an infection by ultra sound. The mare did not get in foal with frozen semen a 2 yrs ago. The mare was taken to the stud farm, stallion was collected and the vet AIed the mare on site with 70ml of raw semen. 15 days after ovulation she was scanned and was in foal but at 30 days not in foal. The owner asked the vet would you do anything different the next time (next spring), she said "no". I use this vet clinic so I'm very concerned if I have a mare that doesn't settle easily.

I want to learn how to do my own mare's cytology, I'm good at AI-ing been doing it for 10 yrs+, I found a source for the cytology brush, I have the microscope, a good microscope camera and several mares to practice on. Do I need special slides (caffine/stain) or anything else? I have a vet friend that will probably help me if needed. How hard can it really be? Any advise?
 

Jos
Board Administrator
Username: Admin

Post Number: 3388
Registered: 10-1999
Posted on Monday, August 22, 2011 - 11:59 am:   Edit Post Delete Post    Move Post (Moderator/Admin Only)

What's a definition of insanity? doing the same thing over and over and expecting a different outcome each time...

If a "reproductive vet" won't do a cytology, they're not a reproductive vet. And if they won't do a cytology, then that indicates that they probably won't be doing other things that should be done reproductively as well.

Find another vet for your reproductive work.

You do not need a special slide for cytology, although frosted slides will give you a slightly better result. If you are only doing a few, stick with the regular slides and save money.

You will need a differential haematology stain kit such as a "Dif-Quick" kit. This consists of 3 different liquids - a fixative and two different stains.
  • Label your slide with indelible marker so you know which side is up;
  • Roll the cytology brush (available from Har-Vet)with the sample in it on the slide;
  • Air-dry it or place it in an incubator or on a slide warmer to speed it up;
  • Dip 3 times in the fixative solution then run the drips back into the bottle;
  • Dip 4 or 5 times in the red stain and run the drips back into the bottle;
  • Dip 4 or 5 times in the dark blue stain and run the drips back into the bottle;
  • Rinse gently for a second or two under a water stream (do not use water under pressure or you will wash off the sample);
  • Wipe the bottom of the slide (the side with no sample on it) with a paper towel;
  • Air dry or put in an incubator or on a slide warmer;
  • Look at it under a microscope.
There are a couple of good websites with cytology pictures, the best of which is probably John Dascanio's site at this location (note that at the time of writing, that link appears not to be working, but hopefully it will be back up in due course). IVIS also has an article by Dascanio here.

It's not hard to do, is quick, and very useful.
 

hedgerow pony farm
Breeding Stock
Username: Lotsofponies

Post Number: 215
Registered: 03-2009
Posted on Tuesday, August 23, 2011 - 12:43 pm:   Edit Post Delete Post    Move Post (Moderator/Admin Only)

Do I need to put the slide in the incubator to dry between dippings? The cells aren't going contaminate the fixative solution when dipping and running back into the bottle are they?

Another question a little off topic:
Can a mare have an infection, get in foal and carry to the 14-15 day twin check then lose it ? I've been told if they have an infection they can't get infoal.
 

Jos
Board Administrator
Username: Admin

Post Number: 3392
Registered: 10-1999
Posted on Tuesday, August 23, 2011 - 03:23 pm:   Edit Post Delete Post    Move Post (Moderator/Admin Only)

No need to dry between dippings. Just go from one to the next. You are not going to contaminate as you used a fixative and even if any cells drop off they will sink to the bottom. You are not growing cultures, so not the same sort of contamination risk.

Mares can indeed lose pregnancies early due to infection, yes. Indeed in rare cases one will see a mare develop placentitis that originated from an infected uterus at the time of breeding, and that is going to occur typically after 2-3 months of pregnancy.
 

Cathy
Breeding Stock
Username: Cathy

Post Number: 374
Registered: 04-2005
Posted on Wednesday, August 24, 2011 - 11:09 pm:   Edit Post Delete Post    Move Post (Moderator/Admin Only)

Running the water over the back of the slide will also rinse it well with less risk of washing off the sample.



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