# Make sample vial

## Get things ready

You need these items:

* 1 × [dilution vial](https://help.sbtinstruments.com/encyclopedia/item-register/vials-flasks-and-liquids/dilution-vial) or 1 × [empty vial](https://help.sbtinstruments.com/encyclopedia/item-register/vials-flasks-and-liquids/empty-vial)
* 1 × [dilution liquid](https://help.sbtinstruments.com/encyclopedia/item-register/vials-flasks-and-liquids/dilution-liquid-5-l)
* 1 × your own [prepared](https://help.sbtinstruments.com/protocols/prepare-your-sample) sample

See also [what-can-i-measure-on](https://help.sbtinstruments.com/encyclopedia/measurements-explained/what-can-i-measure-on "mention") and [](https://help.sbtinstruments.com/protocols/prepare-your-sample "mention").

## Step-by-step

{% stepper %}
{% step %}

### Dilute your sample

The ingredients for your sample vial are simple: Your [prepared](https://help.sbtinstruments.com/protocols/prepare-your-sample) sample mixed with dilution liquid. 

**How much should I dilute my sample?** It depends. See [hit-the-right-concentration](https://help.sbtinstruments.com/protocols/prepare-your-sample/hit-the-right-concentration "mention").

We'll refer to it as your *sample vial* from now on. Put a label on the vial or mark with a lab marker to remember the contents.

{% hint style="success" %}

## Target 0.5–4 million cells/mL for fast results

The more things to count, the faster we are certain about the concentration. Give BactoBox® something to work with and it triggers the *Fast results* feature :fast\_forward:.

Make a dilution series and [**measure the most diluted sample first**](https://help.sbtinstruments.com/protocols/hit-the-right-concentration/dilute-1-100#measure-the-most-diluted-sample-first). Aim for 0.5–4 million cells/mL.
{% endhint %}
{% endstep %}

{% step %}

### Mix sample vial

Mix the sample vial to uniformly suspend the particles within.

There are two options: Mix with vortex mixer or mix by hand.

{% tabs %}
{% tab title="Mix with vortex mixer" %}
{% hint style="success" %}

## We recommend that you use a vortex mixer to mix the sample vial

This is the best way to uniformly suspend the particles within.
{% endhint %}

Place sample vial on vortex mixer and mix for at least **10 seconds**.

<figure><img src="https://4216107837-files.gitbook.io/~/files/v0/b/gitbook-x-prod.appspot.com/o/spaces%2FeYTtd7c0A325muFr5lMW%2Fuploads%2FwPeMZGcuG5MvgAw35Ijs%2Fvortex-mixer--simple-outline--front.png?alt=media&#x26;token=2e746fa7-90c4-44f3-a592-ea9b15432a98" alt="" width="319"><figcaption><p>Vortex mixer</p></figcaption></figure>

{% hint style="success" %}
Ensure that a vortex/cyclone forms in the liquid.
{% endhint %}

{% hint style="info" %}
The vortex mixer is *not* part of the packaging. You must acquire a vortex mixer yourself.
{% endhint %}
{% endtab %}

{% tab title="Mix by shaking" %}
{% hint style="warning" %}

## Only as a last resort

Only shake the sample vial by hand if you don't have a vortex mixer.
{% endhint %}

Shake the sample vial vigorously for at least **30 seconds**.

![](https://4216107837-files.gitbook.io/~/files/v0/b/gitbook-x-prod.appspot.com/o/spaces%2FeYTtd7c0A325muFr5lMW%2Fuploads%2Fsvs6bzT7jIZjMsX6Ujh3%2Fsampling-vial--shaking.jpeg?alt=media\&token=9dd7bddd-1e0f-4ab9-a381-b24c691eec93)
{% endtab %}
{% endtabs %}
{% endstep %}

{% step %}

### That is it!

You now have your own sample vial that you can measure.

{% hint style="warning" %}

## Your sample vial is for *single use* only

Once you run the liquid through BactoBox®, the properties change *significantly*.

**Always discard your sample vial after use!**
{% endhint %}
{% endstep %}
{% endstepper %}


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