Nexcelom Bioscience Cellometer Auto T4, User Manual, Page: 38 / 75

Chapter 7. Sample Preparation
8001009 Cellometer® Auto T4 User Manual Rev C 32
LOADING SAMPLE INTO COUNTING CHAMBER
Cellometer chambered slides consist of two independent counting chambers manufactured to a precisely-
controlled volume. A cell suspension of 20 µL is pipetted into a chamber through one of the induction ports.
1. To prepare the sample for counting, invert the tube containing your sample 10 times, then pipette up and
down 10 times. This will help evenly suspend cells. Do
not shake sample as it may damage cell membranes.
Note: If testing for cell viability ( optional ), stain the sample using a 1:1 ratio with 0.2% trypan blue (or other
chosen viability stain) and mix the stain by pipetting up and down 10 times before performing Step 1, above.
When preparing a sample to test for cell viability, ensure that the trypan blue stock concentration used is 0.2%.
If stock concentration is 0.4%, it is recommended you dilute it with a balanced salt buffer (e.g., PBS) and filter
diluted solution with a 0.2 micron filter before adding stain to your sample.
2. Immediately transfer 20 µL of sample into a counting chamber by pipetting directly onto sample induction
port.
There are two counting chambers per slide; each chamber may be loaded with a different sample.
3. Hold the loaded chamber in the white area and take care to stay away from the clear optical window.
Load the counting chamber by touching the pipette tip to a sample introduction port and then slowly pipette the entire 20 µl
sample at once. Capillary force automatically spreads the cell sample within the counting chamber. The chambers can hold
separate samples as there is no mixing between the two chambers on the slide.