Hello, I am a student studying stem cell engineering.
I am going to stain human stem cell-derived chondrocytes with Alcian Blue staining technique.
I differentiated mesenchymal stem cells using human embryonic stem cells. And chondrocyte differentiation is in progress using mesenchymal stem cells.
In order to identify the chondrocytes that I have differentiated, I am going to use the Alcian Blue staining.
This is the Alcian Blue staining protocol I have.
0. 4% PFA fixation 30min
1. 3% Acetic acid 3min
2. 1% alcian blue solution 30min RT or 15min 37oC
3. washing 3% Acetic acid
4. DW washing
5. mounting
(*alcian blue solution : TMS-010-S, Merck)
I dyed with the above method, but the Alcian Blue dye does not work.
I do 3min with 3% acetic acid and then treat with alcian blue solution for 1 hour. Then it won't be colored.
However, if you do not use 3% acetic acid for 3 min, and proceed with staining with alcian blue solution for 1 hour immediately after fixation, then the dye will be stained.
So, I am very confused.
I am curious as to whether the differentiation into human stem cell-derived chondrocytes is correct.
And I would like to discuss with you about the difference in the dyeing result according to the treatment of 3% acetic acid in the Alcian Blue staining.
I appreciate any help and opinion from us.
Take care and be safe.
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