Always disaggregate

Mini course: A single cell suspension is essential for reliable BactoBox® measurements.

BactoBox® is a flow cytometer. This means that cells are counted individually as they pass through the detector. For the most reliable results, disaggregate cell clumps into a single-cell suspension before measurement.

Single cells can readily enter the measurement channel, while larger clumps of cells must be disaggregated prior to dilution and measurement. Otherwise underestimation of cells may occur.

How to get a single-cell suspension

The best way to disaggregate cells is to use a mechanical method when the cells are still in the growth medium.

Vortex at maximum speed for 1 minute in a small vial like a 2 mL or 5 mL Eppendorf tube.

Master the art of vortexing

Vortexing may look simple. It isn't. Especially not for larger vials. Proper vortexing requires a suitable vortex mixer attachment and a proper hold of the vial.

Slow-motion vortexing of a 15 mL vial. Left: correct grip and correct attachment. Center: too tight grip. Right: wrong attachment.

Correct

The cap is initially held with three fingers to start the motion, after which a single finger applies light downward pressure while allowing the tube to vortex freely.

Make sure that a vortex/cyclone forms.

Incorrect ✘

The vial is held too tightly using a central grip. The liquid bounces slightly, but the vortex does not form.

Incorrect ✘

A 50 mL vortex mixer attachment is used for the 15 mL tube. The vortex does not start even though the grip is correct.

Learn the ropes

This mini course provided you with the essentials on how to disaggregate cell clumps prior to BactoBox® measurements.

Try our first tutorial for a quick, hands-on example on how to disaggregate cell clumps.

Do a simple measurement

Proceed to the next mini-course Hit the right concentration to learn why cultures must be diluted prior to measurements.