Page 1 of 1

Staining of Chromosomes

Posted: Mon Jan 17, 2022 7:15 pm
by MicroscopyLearning
Hello. Recently I have been wondering how to stain the chromosomes of a cell. I have seen multiple sources saying to use the stain Safranin. How would you recommend applying this stain, should it be diluted first and then applied? The safranin I use is a 1 percent aqueous solution. Also, does anyone know how much magnification would be needed to view the chromosomes? Thank you for any advice you can give.

Re: Staining of Chromosomes

Posted: Mon Jan 17, 2022 9:39 pm
by MicroBob
Something like chomosomes in root tips?

Re: Staining of Chromosomes

Posted: Mon Jan 17, 2022 9:44 pm
by Alexander
Chromosones are very much borderline for any light-microscope. I recommend to use fluorescence to visualize them.

Re: Staining of Chromosomes

Posted: Tue Jan 18, 2022 12:13 am
by DrPhoxinus
I have done karyotypes of fish and lizards.
Metaphase is very brief in the cell cycle so to get a reasonable number one has to arrest cells with colchicine.

I used Giemsa stain.

If you are talking about plants you need meristems.

Prepping to see chromosomes can be hard.

Re: Staining of Chromosomes

Posted: Tue Jan 18, 2022 8:04 pm
by Leitzcycler
Chromosomes are difficult to find without using special techniques. The best way to start is to use onion root cells. You may google plenty of procedures. According to my experience the most important thing is to use proper material. You grow roots and only use the very fresh tips. I think any general stain will do although Giemsa is probably one of the best. Chromosomes are of different sizes depending on the organism. Onion chromosomes are rather big and can be well seen with 40x objective.

Good Luck!

Re: Staining of Chromosomes

Posted: Tue Jan 18, 2022 8:16 pm
by DrPhoxinus
Speaking of variation in chromosomes, while in Florida I came across one of these

https://en.m.wikipedia.org/wiki/Amphiuma_pholeter

It has 28 times the DNA as a human and a 10x objective did not have enough field of view for a chromosome spread

Acetocarmine appears good for onions

https://www.vedantu.com/question-answer ... 5b3a0b01f0