Prepare sample

After ~5 hours of incubation, the cell concentration should be sufficient for BactoBox® measurements. The step-by-step guide below describes the sample preparation for all growth curve measurements.

Vortex thoroughly and vigorously

BactoBox® is a flow cytometer and performs best when cells are present as a single-cell suspension.

Because bacterial cells can stick together, thorough disaggregation is essential. One minute of vortexing may seem excessive, but it is necessary to achieve reliable measurements.

Step by step

1

Get a sample of the culture

Make 10 figure-eight swirls to resuspend the culture prior to sampling.

Use a serological pipette to aspirate 2 mL culture and transfer it to a 5 or 15 mL centrifuge vial.

2

Disaggregate clumps of bacteria

Vortex the sample at maximum speed for 1 minute.

Ensure that a proper vortex forms inside the tube during mixing.

3

Set a countdown for the next measurement

Set a 30-minute countdown to remind yourself to collect the next sample for the growth curve.

Summary

You have now collected a sample after ~5 hours of incubation and used vortex mixing to disaggregate bacterial clumps. Proceed to 1:100 dilutions to determine the cell concentration using BactoBox®.