8004708 Cellometer™ Ascend User Manual Rev A 29
Chapter 7. Sample Preparation
This chapter describes preparing Cellometer counting chamber slides and loading counting beads/cell samples into
counting chambers.
PREPARING COUNTING CHAMBER SLIDES
1. For 3-Chamber and 8-Chamber slides, remove protective film from both sides of the slide. Do not touch or write
on the clear optical window areas of the slide.
Note: It may be difficult to peel protective film from the slide. One method is to adhere tape to the top of film
and then pull tape off (with film attached).
2. Place the Cellometer disposable counting chamber slide on a fresh Kimwipe.
3. To prepare multiple samples on the same slide, label individual counting chambers (e.g., #1, #2, etc.) in the
white margin of each chamber. Be careful to not touch the clear optical window of the counting chamber.
Note that Ascend 3-Chamber slides have already been labeled with A, B, and C.
LOADING SAMPLES INTO COUNTING CHAMBERS
Cellometer Ascend 3-Chamber and 8-Chamber consumable slides consist of independent chambers manufactured
to a precisely controlled height. A cell suspension of 20 µL is pipetted into a chamber through its inlet port.
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 to 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 µm filter before adding stain to your sample.
2. Immediately transfer the recommended volume of sample into a counting chamber by pipetting directly onto
its inlet port. For 3-Chamber slides, load 20 µL per chamber. For 8-Chamber slides, load 10 µL per chamber.
3. When handling the slide loaded with sample, be careful to not touch the clear optical windows.
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