staining blood with giemsa

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nigeldodd
Posts: 11
Joined: Fri Jul 03, 2020 10:33 am
Location: cheltenham

Re: staining blood with giemsa

Post by nigeldodd »

thanks, Bill. Your experience is obviously valuable.

"small drop of blood near one end of the slide and then use another slide to make the smear. Place the second slide at about a 30 degree angle to the smear slide with the edge just ahead of the drop. Then back up the second slide until it just touches the drop of blood and smoothly push the spreader slide forward." Yes, this is what I tried to do. I can understand that it is desirable to have a smear only one cell thick. My smear, when looked at without a microscope, is roughly comma shaped, but you say it is still too thick.

Could you please explain what is meant by "fixing" in your comment "methanol in the stain fixes the cells". What, actually, is "fixing"? In photography the term is used to describe the removal of unexposed silver, but of course this is not the same.

I am also struggling to understand how to precisely measure ph, which seems necessary to make a buffer solution.

And the main problem is how to obtain small quantities of these chemicals.

I have CLL and it would be useful and insightful to see the white blood cells.

thanks again

Ichthyophthirius
Posts: 1094
Joined: Thu Mar 07, 2013 5:24 am

Re: staining blood with giemsa

Post by Ichthyophthirius »

Hi,

[If you can't get the buffer, use the Evian method (link above).]

I'm very sorry to hear that. As you can see, even very basic diagnostics can be difficult and are highly regulated to ensure reproducible results. I strongly advice against using any home diagnostics as a basis for treatment decisions and to go through your GP. My advice was really just meant for blood smears made for curiosity.

All the best,

Ichty

nigeldodd
Posts: 11
Joined: Fri Jul 03, 2020 10:33 am
Location: cheltenham

Re: staining blood with giemsa

Post by nigeldodd »

Please rest assured that I am not using home diagnostics. I am seeing a team of haematologists and am not having treatment yet.

However I am naturally curious and have done a great deal of research around the subject. My PhD is in physics but I am reasonably quick to assimilate new subjects.

But it is proving tricky to get to see the white blood cells. I am hugely appreciative of all the advice I have received in this forum. When time permits, I will do some more smears.

billjanes1
Posts: 88
Joined: Fri Dec 30, 2016 1:59 pm
Location: Lake Forest, IL, USA

Re: staining blood with giemsa

Post by billjanes1 »

Sorry to hear that you have CLL.

Fixation preserves and stabilizes the specimen prior to processing. For blood and bone marrow smears absolute methanol is generally used. For tissue biopsies formaldehyde is often used.

For giemsa staining the smear is fixed in methanol and then placed in diluted giemsa stain for a more prolonged time. See this link. Also note the quick giemsa method which is similar to that used for Wright staining.

https://www.sigmaaldrich.com/content/da ... 1/gs10.pdf

Wright stain contains methanol and no separate fixation is needed. The stain is applied full strength to the slide and the methanol in the stain fixes the smear. Distilled water or buffer is applied and staining proceeds. See this link. Note the comment that the slide must be absolutely clean or the stain won't work.

https://www.polysciences.com/skin/front ... df/801.pdf

Bill

nigeldodd
Posts: 11
Joined: Fri Jul 03, 2020 10:33 am
Location: cheltenham

Re: staining blood with giemsa

Post by nigeldodd »

I have been cleaning the slides in the dishwasher. Perhaps it leaves a residue of wetting agent?

Is there a way to test my Giemsa to make sure it has not been contaminated?

thanks again.

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