To set up QTracker (specifically the Invitrogen Qtracker Cell Labeling Kits Go to product viewer dialog for this item.
by Thermo Fisher Scientific), you need to follow a precise 5-step laboratory protocol. This process safely delivers fluorescent nanocrystals into the cytoplasm of live cells for long-term tracing. 🧪 The 5-Step QTracker Setup Protocol
Follow these five steps to properly mix the reagents, label your cells, and prepare them for imaging: 1. Pre-Mix the Components
Combine exactly 1 µL of Qtracker Component A and 1 µL of Component B in a sterile 1.5 mL microcentrifuge tube.
Incubate this concentrated mixture for 5 minutes at room temperature.
Note: Always proceed immediately to the next step once the 5 minutes are up. 2. Prepare the Solution
Add 0.2 mL of fresh, complete growth medium directly into the microcentrifuge tube containing your mixed components.
Vortex the tube thoroughly for 30 seconds to ensure the labeling solution is completely homogenous. 3. Add the Cells
Introduce 1 × 10⁶ cells into the prepared labeling solution.
If you are working with a cell suspension, it is best to pull this aliquot from a stock concentration of approximately 1 × 10⁷ cells/mL. 4. Incubate the Sample
Place your cell and solution mixture into a laboratory incubator. Incubate the sample at 37ºC for 45 to 60 minutes.
Tip: If you are using an adherent cell line, you can adjust this step by applying the solution directly to cells grown on chambered coverglass or sterile coverslips. 5. Wash and Visualize
Wash the labeled cells twice using fresh, complete growth medium to remove any un-incorporated nanocrystals.
Image or analyze the live cells immediately using fluorescence microscopy or flow cytometry equipped with the appropriate excitation/emission filters. 💡 Key Technical Tips for Best Results
Target Concentrations: The standard, final working concentration for an optimal Qtracker label ranges from 2 nM to 15 nM, depending heavily on your specific cell line.
No Cross-Contamination: Once inside the cytoplasm, the intense fluorescent labels will track through several cellular generations, but they will not transfer to adjacent, un-labeled cells in a mixed population. If you are working with a specific cell type, let me know: What cell line are you currently using?
Are you planning to use fluorescence microscopy or flow cytometry for your analysis?
I can give you the exact filter recommendations and protocol adjustments for your specific experiment! Qtracker® Cell Labeling Kits – Thermo Fisher Scientific
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