> 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/mpd/workflows/cross-check/calculations-for-plating.md).

# Calculate dilutions

For the cross-check, you will also need the plate count results. The cell concentration provided by BactoBox® measurements gives you a good estimate of how much to dilute the culture. We refer to this as the *assumed optimal dilution*.

We include the vials next to the expected optimal dilution to account for potential differences between BactoBox and plate counts. We refer to this as [Plate smarter](/mpd/workflows/plate-smarter.md#bracketing-dilutions).

The strategy for hitting the optimal dilutions are detailed below. We assume that 1 mL is plated. More information is available in [Plate smarter](/mpd/workflows/plate-smarter.md).

<figure><img src="/files/3RLZTHa1Ymd4zuKgBiUO" alt=""><figcaption><p>Dilution series for plate counts. The 15 mL centrifuge vial has a concentration of 1×10<sup>10</sup> cells/mL. A 1:10 dilution is prepared by transferring 1 mL sample to 9 mL buffered peptone water (or similar). The lowest numbers shows the cells/mL for the individual vials. Eight 1:10 dilutions are needed to dilute to 100 cells/mL. We recommend <a data-mention href="/pages/cKM6SEfU02gZx9C7mkFS#bracketing-dilutions">/pages/cKM6SEfU02gZx9C7mkFS#bracketing-dilutions</a>. Use the cyan bracket when plating 1 mL. Use the magenta bracket when plating 0.1 mL.</p></figcaption></figure>

## Step by step

{% stepper %}
{% step %}

### Write concentration with thousand separators

Write <kbd>cells/mL × dil.</kbd> with thousand separators.

If the cell concentration was 1×10<sup>10</sup> cells/mL, you would write <kbd>10 000 000 000</kbd> .
{% endstep %}

{% step %}

### Determine the assumed optimal dilution

Determine how many zeros to remove to reach 25–250 cells/mL.

In the above example you remove eight zeros from <kbd>10 000 000 000</kbd> to get <kbd>100</kbd>.

In this example it requires <kbd>eight</kbd> 1:10 dilutions to dilute to 25 - 250 cells/mL. <kbd>10</kbd><sup><kbd>-8<kbd></sup> is therefore the assumed optimal dilution when you plate 1 mL.
{% endstep %}

{% step %}

### Bracketing: Include the dilutions next to the optimal dilution

Include the two vials next to the assumed optimal dilution for plating. This "bracketing" approach is a precaution for potential differences between BactoBox® and plate counts.

In the illustrated example we would include the 10<sup>-7</sup> and 10<sup>-9</sup> vial. In this example we would plate the 10<sup>-7</sup>, 10<sup>-8</sup>, and 10<sup>-9</sup> vials (<mark style="color:cyan;">cyan bracket</mark>).
{% endstep %}
{% endstepper %}

{% hint style="info" %}

## Use one dilution less if you are only plating 0.1 mL

* The above calculations assume you use 1 mL for plating. This corresponds to the <mark style="color:cyan;">cyan bracket</mark> in the illustrated example.
* Use one dilution less if you are using the spread plate method with only 0.1 mL. This corresponds to the <mark style="color:purple;background-color:violet;">magenta bracket</mark> in the above illustration.
  {% endhint %}

## Summary

You have now determined which dilutions to use for plating. Next step is [Plate dilutions](/mpd/workflows/cross-check/plate-dilutions.md)<br>


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