some commercially available fluorescent dye like  CellTracker™ Probes, but they need harvested samples for slide preparation but for cell tracking i have only one animal to track cells 30 and 60 days.

Could you try to mark your cells with GFP before seeding?

There is the vibrant red from molecular probe that is a usefull tracker, fixation resistant!!

yes i am in need of any fluorescent labeling fluorochrome for cell membrane protein its may be osteocalcin.

thanks for your kind suggestion, vibrant red is ideal tool for nuclear stain but i am in need of cell membrane staining or whole cell tracking by whole cell staining, but now i am trying PKH26 red fluorescent dye (Sigma Aldrich).

Than you can may be try LDL DiI, that bounds the cell mambrane beeing lipophilic dye.

I would suggest DiI, based on my experience (different cell types), it should work good enough. 

Thanks for your kind suggestions, I used PKH26 on my cells and it gave good fluorescent in compare to unstained cells. I  checked it from P1 stage but it will go through P4 or P5 stage in-vitro. i will try LDL DiI stain later on.

osteoblasts on the scaffold I would go for smartflare specific to osteogenic markers such as RUNX2 (CBFA1) or SPP1 (osteopontin) as they will bind to the upregulated transcript of RUNX2 or SPP1 (if human) and as they are already transcribing these genes. also remember decellularised bone will be fluorescent. we seeded hMSC in to chick femurs and had to overcome the auto-fluorescent nature of the bone by decalsification.

Dear Netrapal,

In my PhD research I often used the fluorescent dye call CDIL traker, with this probe you can track the viable cells in green (without killing the cells) and if you add IP (propidium iodid)you can reveal in red the dying cells.

Best,

Philippe Bouaziz