EVOS™ M7000 Imaging System Quick Reference

Introduction

This document provides a quick reference for operating the EVOS™ M7000 Imaging System for fluorescence and transmitted light applications. It covers essential steps for capturing images, adjusting settings, and managing image display.

1. Capture Images

Turn on the EVOS™ M7000 Imaging System:

1. Turn the instrument power switch to ON.
2. Turn the computer and monitor ON.
3. Click the M7000 icon to start the EVOST™ M7000 software.
4. Ensure the Capture tab is displayed.

Select sample vessel:

1. Place the vessel containing your sample on the stage using the appropriate vessel holder.
2. On the Capture tab, click the Vessel button to open the Vessel Selection dialog.
3. Select the Vessel category, Holder, and Vessel type.
4. Click Done.

Select objective and light source:

1. Click the desired Objective button (e.g., 4x, 10x, 20x, 40x).
2. Select the desired Light source (e.g., DAPI, GFP, RFP, CY5, Trans) for which you wish to adjust brightness and set focus.

2. Focus on the Sample

Autofocus:

• Click Autofocus to run the autofocus procedure for the current channel and sample.
• Alternatively, use the Coarse and Fine focus sliders to manually find the best focal position.

Optional: Set Z-Offsets:

• Verify that the Lock Z-Offsets option is checked in the Advanced focus settings window.
• Select the Objective and Light source.
• Click the Light button to enter Live mode and focus manually or using Autofocus.
• After finding the optimal focus in the first channel, uncheck Lock Z-Offsets in Advanced focus settings.
• Select the next Light source and repeat the focus procedure.
• Repeat for all desired channels.
• When finished, check the Lock Z-Offsets option again.

3. Set Brightness and Camera Options

Click the Brightness and camera settings button to expand the controls.

• Mode: Select Simple (single Brightness parameter) or Actual (adjust Light, Exposure, Gain individually).
• Camera: Select Mono (for fluorescence and transmitted light) or Color (for brightfield only).
• Phase options: Available for transmitted light channels.

Click the Brightness and camera settings button again to collapse the controls.

4. Adjust Brightness

Click the Light button to turn on the excitation light and enter Live mode.

• Adjust brightness using the Brightness controls (Simple mode) or Light, Exposure, and Gain controls (Actual mode).
• Note for best results: Optimize parameters for searching (increase Gain, decrease Exposure) and capturing (decrease Gain, increase Exposure). For time-lapse imaging, increase Gain and Exposure, and decrease Light intensity to reduce photobleaching.

5. Set Image Display Options

Choose channels to display by selecting the appropriate channel checkbox on the Light source button (e.g., DAPI, RFP, Trans).

• Click the Image Display settings button to expand the controls.
• Adjust Brightness, Contrast, and Gamma settings using the sliders for each selected channel.
• Unselect a checkbox to remove a channel from the Viewing Area.
• Click the Image color display button to display samples in pseudo-color.

Click the Image Display settings button again to collapse the controls.

Note: Adjustments to Image display settings only affect the viewing, not the capture.

6. Select Field and Capture Images

• In Field View or Live mode, position the capture crosshair over the region of interest.
• Use the Jog Control or navigation arrows to move the stage.
• Click the Light button to exit Live mode if necessary.
• Click Capture to acquire an image of the selected field using the current settings. A thumbnail of the captured image is displayed.
• To capture the same field in another channel, select the desired channel and click Capture.
• To capture a different field, position the crosshair over the new region of interest and click Capture.
• To capture a field using a different magnification, select the desired Objective and repeat the capture procedure.
• To capture a selected field in multiple channels simultaneously, check the circular checkboxes on the corresponding light source buttons and click Capture Channels.

7. Save Captured Images

1. When finished capturing images, click Save... to open the Save dialog.
2. Select captured fields to save: Choose from Currently selected field, All newly captured fields, or All captured fields.
3. Select save location: Click Browse to navigate to the desired folder. It is recommended to save to an external hard drive.
4. Select file types to save: Check the desired checkboxes for Raw images (recommended for analysis) and Displayed images. Configure file formats (TIF, PNG, etc.), color options (Grayscale, Pseudocolor), and grid options as needed.
5. Click Save to save the images in the designated folder.

Documentation and Support

For the latest services and support information, visit thermofisher.com/support.

At the website, you can access FAQs, submit questions to Technical Support, search for user documents, SDSs, application notes, and download software updates.

Safety Data Sheets (SDSs) are available at thermofisher.com/support.

Limited Product Warranty information can be found on the Life Technologies website.

For Research Use Only. Not for use in diagnostic procedures.

Manufacturer: Life Technologies Corporation, 22025 20th Ave SE St #100, Bothell, WA 98021.

All trademarks are the property of Thermo Fisher Scientific and its subsidiaries.