Giemsa stain of thin blood smears - pale / faint
Posted: Tue Apr 23, 2024 9:54 pm
I have a question about the Giemsa staining of thin blood smears
I've been doing this for a few months now mainly studying red blood cells, white blood cells, platelets etc
The main problem I'm having is I'm finding it difficult to get the stain and dark enough especially to darkly stain the red blood cells using Giemsa stain
I'm currently making my own Giemsa stain solution from Giemsa powder from a microscopy stain supplier - and using about 1 g of Giemsa stain powder in 100 mL of pure methanol heating it and agitating gentility with glass beads for around 3-4hrs at 50-60 deg C then cooling and filtering it and using that solution neat in a Koplin Jar – immersing the methanol fixed slides for around 10minutes ( longer doesn’t seem to help ) before rinsing briefly in water ( if I rinse for more than around 3 seconds I get even less contrast – but any less than this and I get lots of stain artefacts).
staining is sort of okay but I just need ideally more contrast between the red blood cells and the background.
When viewed at 1000x under oil there is just a lack of contrast between the red blood cells and the background – and similar with white blood cells – white blood cells are stained kind of okay, the nucleus is visible - but again, they could be darker so it seems to be a general issue with the stain not being dark enough so am I hope you can help with that.
Any suggestions for increasing contrast especially in the red blood cells and any contents inside them
i will attach an image as an example - (the colour balance is doing something a bit off - but its not affecting the contrast much - just making the entire image more yellow than my last camera)
Much appreciated !
I've been doing this for a few months now mainly studying red blood cells, white blood cells, platelets etc
The main problem I'm having is I'm finding it difficult to get the stain and dark enough especially to darkly stain the red blood cells using Giemsa stain
I'm currently making my own Giemsa stain solution from Giemsa powder from a microscopy stain supplier - and using about 1 g of Giemsa stain powder in 100 mL of pure methanol heating it and agitating gentility with glass beads for around 3-4hrs at 50-60 deg C then cooling and filtering it and using that solution neat in a Koplin Jar – immersing the methanol fixed slides for around 10minutes ( longer doesn’t seem to help ) before rinsing briefly in water ( if I rinse for more than around 3 seconds I get even less contrast – but any less than this and I get lots of stain artefacts).
staining is sort of okay but I just need ideally more contrast between the red blood cells and the background.
When viewed at 1000x under oil there is just a lack of contrast between the red blood cells and the background – and similar with white blood cells – white blood cells are stained kind of okay, the nucleus is visible - but again, they could be darker so it seems to be a general issue with the stain not being dark enough so am I hope you can help with that.
Any suggestions for increasing contrast especially in the red blood cells and any contents inside them
i will attach an image as an example - (the colour balance is doing something a bit off - but its not affecting the contrast much - just making the entire image more yellow than my last camera)
Much appreciated !