> 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/simple-measurement.md). # 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[^1] 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[^1] 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.

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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.** {% endcolumn %} {% column %}

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

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A 50 mL vortex mixer attachment is used for the 15 mL tube. The vortex does not start even though the grip is correct. {% endcolumn %} {% endcolumns %} ## Learn the ropes This mini course provided you with the essentials on how to disaggregate[^1] cell clumps prior to BactoBox® measurements. Try our first tutorial for a quick, hands-on example on how to disaggregate[^1] cell clumps. {% content-ref url="/pages/yOiWxVnPgOuX7cibH9R9" %} [Do a simple measurement](/not-in-the-new-structure/measure-vitroid-tm-disc.md) {% endcontent-ref %} Proceed to the next mini-course [Hit the right concentration](/more/how-to-guides/hit-the-right-concentration.md) to learn why cultures must be diluted prior to measurements. [^1]: Break up clumps or chains of cells --- # Agent Instructions This documentation is published with GitBook. GitBook is the documentation platform designed so that both humans and AI agents can read, navigate, and reason over technical content effectively. Learn more at gitbook.com. ## Querying This Documentation If you need additional information that is not directly available in this page, you can query the documentation by asking a question. Perform an HTTP GET request on the following URL with the `ask` and `goal` query parameters: ``` GET https://help.sbtinstruments.com/not-in-the-new-structure/simple-measurement.md?ask=&goal= ``` `ask` is the immediate question: it should be specific, self-contained, and written in natural language. `goal` is what the user is ultimately trying to achieve, the reason they need the answer. Sharing it helps GitBook give you a better, more relevant answer. A goal is most helpful when it describes the outcome the user wants rather than restating the question. For example, with `ask=how do I create an API token`, a goal like `automate deployments from our CI pipeline` lets GitBook tailor the answer to that use case. The response will contain a direct answer to the question and relevant excerpts and sources from the documentation. Use this mechanism when the answer is not explicitly present in the current page, you need clarification or additional context, or you want to retrieve related documentation sections.