Summary

In this tutorial you learned how to prepare a dense bacterial culture for BactoBox® measurements. After thorough disaggregation and 1:10,000 dilution, you got a fast result.

Calculate cell concentration of non-diluted suspension

The BactoBox® result was for the diluted sample. Always remember to multiply the obtained result with the dilution factor to get the concentration of the non-diluted suspension.

BactoBox® dilution	BactoBox® concentration	Non-diluted suspension
10 000	540 000 cells/mL	10 000 × 540 000 cells/mL = 5.4 × 109 cells/mL

BactoBox® skills

Congratulations! After successfully completing this tutorial you have acquired the following skills:

Disaggregrate bacterial clumps by vortexing

Make a 1:10 000 dilution of a dense bacterial culture and measure on BactoBox®

Calculate cell concentrations of the non-diluted bacterial suspension

Proper disaggregation is extremely important

Disaggregation of cell clumps is important for any bacterial enumeration method. But it is especially important for BactoBox® measurements. The measurement channel in the flow cell will only accommodate objects smaller than 5 µm spherical diameter. Larger objects will stay in the bypass channel. Clumpy, bacterial suspensions will be severely underestimated on BactoBox® unless proper disaggregation is done.

Vortex works well for most liquid cultures of bacteria, but sometimes you need more advanced sample workup to Break up clumps and chains.

Next steps

This, second tutorial was a simple introduction on how to measure a concentrated bacterial culture on BactoBox®.

We have a third and last tutorial on cell measurements for you: Track a growth curve. In this tutorial you will inoculate LB medium using a Vitroids™ disc (or an in-house E. coli cryo stock). After a few hours of incubation you will use BactoBox® to track a beautiful growth curve.

We have a third and last tutorial on cell measurements for you: Track a growth curve. In this tutorial you will inoculate LB medium using a Vitroids™ disc (or an in-house E. coli cryo stock). After a few hours of incubation you will use BactoBox® to track a beautiful growth curve.

We will soon have a third and last tutorial on cell measurements for you. In this tutorial you will inoculate LB medium using a Vitroids™ disc (or an in-house E. coli cryo stock). After a few hours of incubation you will use BactoBox® to track a beautiful growth curve.