A. Several things influence image speed acquisition:

1- Number of Channels: to achieve the fastest acquisition, image with one channel.

2- Exposure time: Make sure that you are using the minimum exposure time. If the signal is bad, try to use higher intensity and more sensitive camera (axiocam 702).

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Axiocam702: 0.1ms − 60 s.

3-Number of steps of a Z-stack: the less is better.

4- ROI: the smaller the ROI in Y-axis -the faster you will be able to image.

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5- Camera: CD7 has two cameras, Axiocam702 is faster however Axiocam506 has a smaller exposure time.

• CMOS Axiocam702: Up to 128 fps at full resolution and up to 1000 fps @ 1024 × 128 pixels.

• CCD Axiocam506: Binning Pixel: 1x1, Count (H x V): 2752 x 2208, Mode Mono, 20 FPS @ 1 ms.

6- Pupil component: If you want to do past imaging in the transmitted light channel (Oblique), make sure that the system doesn't waste time taking in and out the “pupil” component; it should be in.

In the “Imaging Setup” tab, go to advanced, go to “After oblique:…” and unclick the box “include in this setting”-red arrow.

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B. How to set up a fast acquisition:

Without tiles setup/

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single position:

Once you set everything up (channels, position, focus, etc..)

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**Once “Use Burst Mode if Possible“ is activated, the system buffers acquisition data in the main memory. Burst Mode enables the recording of the fastest framerates, which the used camera hardware can achieve. This mode supports only single-channel time-lapse acquisition.

 Start the experiament.

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With tiles setup / Multiple positions:

Once you set everything up (channels, position, focus, etc..)

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