Nuclei Counting with Luna FL cell counter

**protocol is provided by one Luna FL user lab at UC Davis**

1. Prepare and load your sample.
   1. Mix 20 µL sample of your cell suspension with 20 µL of 1X Diamond Nucleic Acid dye.
   1. Place a clean Luna Reusable Slide Covers carefully on top of the Luna Reusable Slide chamber.
   1. Place the pipette into the space between slide and coverslip at the edge of the chamber. Load 10 µL mixture into the chamber of Luna Reusable Slide.
2. Turn on the Luna-Fl and insert the slide.
   1. Insert the slide completely into the slide port of the counter.
3. Count cells.
   1. Select “Fluorescence cell counting” mode.
   1. Go to “Protocol” tab and select MFP DF2 on the list of protocol and click “Load”.
      1. Dilution factor = 2.00
      1. Minimum cell size = 5
      1. Maximum cell size = 60
      1. Green fluorescence threshold = 7
      1. Red fluorescence threshold = 7
   1. Use the focus knob to focus on the sample.
   1. Press “Bright field” to switch the fluorescence preview mode. Adjust the level of exposure.
   1. Make sure that the cells have settled down before pressing count.
   1. Press “Count” to start cell counting.
   1. Pipette 10 µL of the mixture and conduct 3 readings. Repeat this process two more times for a total of 9 replicate samples. Calculate the average count.
   1. The automated cell counter provides the concentration of your cell suspension in cells/mL. To convert to cells/µL, simply divide the given concentration by 1000.

contact info@wisbiomed.com for any questions or request complete protocol with dye pack preparation protocol.