细胞世界

2-10 million cells/60-mm petri-dish. 60-200 ng/ml sRANKL.

Osteoclasts appeared from day 3 on.

The purpose of countstain the cells with Hematoxylin is to see nuclei. Nuclei are blue labelled by Hematoxylin, not the color what you mentioned. Your are right, countstaining is not necessary for there is no problem to see nuclei without this step.

What is the cat# of Sigma kit you are using? I used to use 386/7-A but I do not like it.

I made sRANKL by myself so does not matter how big the cell culture vehicle used. However,if you purchase RANKL you should use 8-well chambered slides, of course depends on what is the purpose of your experiments. For instance, if you want to extract RNA from the cells, you have to use dishes otherwise you would not have enough products. If you want to see osteoclasteogenesis, I think either 8-well chambered slides or 48-well plate will do.

For 60-mm Petri-dishes, 4 ml medium/dish.

I know commercial RANKL is very expensive, around 180USD/10ug.