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TutorialBeginner
Do a simple measurement
This tutorial is for your first bacterial measurement on BactoBox®. The tutorial is simple, fast, and predictable. You will learn how to prepare bacterial suspensions for BactoBox® measurements.
For this tutorial, the success criterion is to get a BactoBox® measurement within 30 000 to 5 000 000 cells/mL.
The below table lists the overall considerations for this tutorial. We use similar tables for our microbial process development Workflows so you know if the workflow is right for you and what time to set aside.
You can complete this tutorial as a 🎒Beginner to BactoBox® measurements.
For this tutorial you need to procure Escherichia coli WDCM 00013 Vitroids™ ATCC® 25922 from Sigma/Merck. Lead time is usually 1-5 days. Price is ~200 USD including shipping. 10 vials are included in each shipment. Concentration per disc is stated as 50,000-150,000 CFU mean value range.
The discs are convenient for this tutorial as they dissolve easily in water and have a well-defined concentration of bacteria within the working range of BactoBox® measurements.
A soluble Vitroid™ disc containing 50,000–150,000 CFU of E. coli ATCC® 25922 .
How to get a successful measurement on BactoBox®
Three criteria are essential to get a reliable cell concentration on BactoBox®. Below is the checklist.
The sample should ideally be a single-cell suspension, i.e. without cell clumps.
Conductivity must be within 1,500–2,200 µS/cm.
Concentration of the diluted sample must be between 30,000 - 5,000,000 objects/mL
The most important learning of this tutorial is how to get a single-cell suspension. The below video explains why large clumps must be disaggregated before the individual cells can enter the measurement channel.
Overview
In this tutorial we will demonstrate how to solubilize the disc and get the bacteria ready for BactoBox® analyses. The overall steps in the tutorial are given below.
