Please see these 2 videos:
https://www.bing.com/videos/search?q=counting+by+hemocytometer&view=detail&mid=B7B396F5EDD3CDD17379B7B396F5EDD3CDD17379&FORM=VIRE
https://www.bing.com/videos/search?q=counting+by+hemocytometer&view=detail&mid=B0589E32ED838AC039EBB0589E32ED838AC039EB&FORM=VIRE
Thnx so much but i want to know how to prepare the bone marrow sample from the rat to count
The procedure I followed a while ago:
Cut hind limbs of mouse. Clean fat, muscles, tendon very well. Keep on ice on sterile dish. Rinse the limbs once in PBS. Cut tibia and femur at the ends with a scissors or scalpel.
Flush tibia and femur with 10ml syringe and 25gz needle with staining buffer. Swirl the contents several times with syringe to make it single cell suspension. Keep the dish on ice. Place an iron sieve (autoclaved sieve) on a 50ml tube. Transfer your content into the tube. They will get filtered through the sieve. Place the tube on ice. You can use your bone marrow cells as you have planned for Facs sorting or whatever reason.
Composition of Staining buffer: HBSS GIBCO 14025 with Ca and Mg
10mM Hepes
2% FCS
0.5%bovine serum albumin
Filter
If you have any question, let me know
Good luck
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