# Photomicrography
[EcoEvoDevoLab](https://hackmd.io/@EcoEvoDevoLab/AngeliniLab)

Updated 7 September 2021

This protocol describes how to use Motic camera and software in the Angelini Lab at Colby College.

Before you begin, be sure you have a good way to arrange your specimens. For live insects, be sure you can sedate them (with CO~2~ or cold); for other specimens be sure you know how to orient the specimen towards the camera. Be sure you have adequate lighting. It is also helpful to log into the computer under your own username, and be able to connect to the fileserver.

:::info
For more detail on photography in general and tips on exposure and composition, check out the [tutorial on macrophotography](https://hackmd.io/@aphanotus/macrophotography).
:::

## Imaging

- Be sure the level on the trinocular microscope is set to allow light to the camera. (If you look through the eyepeices, only one should show you the stage. The other should be black.)
- Open both **Motic Images Plus 3.0** and **Motic Images Devices**. (Motic Images Plus is what we use to capture the images, but Motic Images Devices must also be running in the background.)
- In **Motic Images Devices**, under **Basic Adjustment**, select **Motic 10.0P** as the Video Device.
- Click **Open**. (If this doesn't work, click Close, then try Open again. The screen should turn from gray to black or to a live image from the camera. If nothing shows up, unplug and then replug the black cable from the camera to the computer. Then try these steps again.)
- Under **Resolution**, select the maximum value.
- Be sure the lightings are on.
- Set the **exposure**. (Typically, don't adjust gain, offset, enhance, gamma)

:::warning
Often the **Auto** option for Exposure does not produce good results. It can be helpful to start by clicking Auto, but then experiment with setting a manual value, typically higher, to produce a well-exposed specimen. The background can be over-exposed.
:::

- If the color looks odd, aim at a background, white area of the pad and click **White Balance**.
- In **Motic Images Plus 3.0**, click on **Live Preview**  in the left-hand toolbar to see a live image of your view area. (When you first open the app, you may need to click (also located in the left hand-toolbar) in order for the live preview option to become available.
- Right click on the view panel, and select **Fit to Window**. The panel will now show the entire view field.
- Adjust the specimen in the view field. For most insects our lab studies, it is ideal to have a millimeter-scale ruler at the right hand edge of the view field, and to have the insect oriented with its anterior to the left. Center the insect, and keep all appendages in the view field.
- **Focus** on the anatomical features that are most important to your study (for example, the wing venation, or beak).


- To capture an image, select **File/Save As** from the top menu.
- Set the file format to **jpeg image file**

:::warning
You must select the file type every time you save an image!
:::

- Name the file in a way that encodes important information for your study. Be consistent.
- Click **Save**. The live view will be frozen in the frame that was saved.
- Click on **Live Preview** again. [](https://i.imgur.com/yx9OtNA.jpg) and follow the steps above to capture another image.

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[EcoEvoDevo Lab](https://hackmd.io/@EcoEvoDevoLab/AngeliniLab)