> For the complete documentation index, see [llms.txt](https://help.sbtinstruments.com/llms.txt). Markdown versions of documentation pages are available by appending `.md` to page URLs; this page is available as [Markdown](https://help.sbtinstruments.com/not-in-the-new-structure/measure-overnight-culture/disaggregate-bacteria.md).
# Prepare sample
After \~16 hours of incubation, the overnight culture is at a high cell concentration and we can use it to demonstrate BactoBox® measurements on concentrated bacterial suspensions.
{% hint style="success" icon="bacteria" %}
## Vortex thoroughly and vigorously
BactoBox is a flow cytometer and works best when cells are present in a single-cell suspension.
Cells stick together and you need thorough disaggregation[^1] to prepare a single-cell suspension.
One minute of vortexing may sound excessive, but trust us. You need it!
{% endhint %}
## Step by step
{% stepper %}
{% step %}
### Get a sample of the culture
Make figure-eight swirls to suspend the culture
Use a serological pipette to aspirate 2 mL culture.
Transfer to a 5 or 15 mL centrifuge vial.
{% endstep %}
{% step %}
### Disaggregate clumps of bacteria
Vortex the sample 1 minute at max speed.
Ensure that a proper vortex is seen inside the tube when vortexing.
{% endstep %}
{% endstepper %}
## Summary
You retrieved a sample of the overnight culture and used vortex mixing to disaggregate[^1] clumps of bacteria. Proceed to [Dilute and measure](/not-in-the-new-structure/measure-overnight-culture/dilute-and-measure.md) to determine cell concentration with BactoBox®.
[^1]: Break up clumps or chains of cells
---
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