When I label with Qtracker cell labeling reagents, I get a punctate label pattern. How do I make it more uniform?
Qtracker cell labeling reagents are taken up by the cell through endocytosis and sequestered in endosomes. This gives the label a punctate or vesicular appearance. This is normal. There is nothing that can be done to make it appear uniform throughout the cytoplasm.
How long after injection should I start imaging my mouse?
The imaging time course varies with the nature of the injected agent. Vascular tracers are visible in the blood vessels immediately after injection and may be imaged for several hours. Conjugated whole IgG antibodies reach their targets within a few hours of injection and may be imaged for several days.
What happens to unconjugated dye in the mouse after small animal in vivo imaging?
The dye will be eliminated via the bladder. The bladder signal is detectable within ~3 minutes of IV injection of the dye and clearance with ~30 minutes.
What amount of Qtracker non-targeted reagent should I inject to image vasculature?
A recommended starting dosage is 25-50 µL of Qtracker reagent diluted to the desired injection volume with PBS or normal saline. Qtracker reagent should be diluted immediately prior to injection. DO NOT STORE DILUTED. You will need to determine the optimal dosage for your experimental models.
I want to track my cells with a nucleic acid stain, like DAPI or Hoechst dye. Do you recommend this?
This is not recommended. When these stains bind to DNA and RNA, they may affect the normal function of the nucleic acids, disrupting transcription, as well as replication. Other reagents, such as CellTracker dyes or Qtracker reagents are more optimized for tracking without disrupting normal activity. If a nuclear label is still desired, though, and the cells are mammalian and non-hematopoietic, CellLight nuclear reagents can transiently transfect cells to express GFP or RFP on a nuclear-expressing protein for up to several days without affecting function.
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