BakingTray
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BakingTray Documentation
Getting Started
Hardware requirements
Known Issues
Installation
Stitching data
Users
Introduction
Sample preparation
User Guide
Excitation choices
Selecting a resolution
Problems and solutions
Data structure
Legacy User Guide
Starting BakingTray
Step 1: Setting imaging parameters
Step 2: Preparing the sample
Step 3: Selecting the imaging area
Step 4: Starting the acquisition
Step 5: Concluding the acquisition
Setting up checklist
Resuming an acquisition
autoROI
Imaging multiple brains at once
Developers
Developers
FAQ
Gallery
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Setting up checklist

Common steps

* Select Sample -> New from the menu.
* Turn on the laser and open the shutter
* Embed the sample
* Load sample into microscope.
* Set sample directory and sample name.
* Set imaging parameters in main BakingTray window.
* Set channels to acquire in ScanImage CHANNELS window.
* Raise sample and set blade position.
* Take the first slice: ensure blade ends up about 3 mm beyond the edge of the agar.
* Trim off excess agar and approach imaging depth.
* Hit the Auto-Trim button to go from thicker trimming section to the final slice thickness and speed used for imaging.
* Center sample under objective. Open laser shutter. Turn off room lights.
* Find the sample surface, take a slice, then re-measure surface to check for stability.

autoROI

* Find sample ventral midline whilst Imaging. Press "Set Ventral Midline" to set initial imaging area.
* Take a preview scan to ensure that samples don't enter the green border area. This can be at lower power for now. If necessary, increase imaged area. You don't need to re-take the preview scan right now.
* Increase frame averaging to tweak bidirectional scanning. (You can middle-click on the preview image to move the stages to that location in the sample)
* Set laser power and power increase with depth at an area without too much white matter.
* Reset frame averaging and *take another preview* at the final power and PMT settings. Hit AutoThresh.
* BAKE
* After acquisition you should crop the stitched images from even single samples or you will end up with larger datasets than you expect.

Non-autoROI

* Find sample ventral midline and use this to set initial imaging area.
* Take a preview scan to ensure that samples are correctly framed. Re-frame as needed.
* Increase frame averaging to tweak bidirectional scanning. (You can middle-click on the preview image to move the stages to that location in the sample).
* Set laser power and power increase with depth at an area without too much white matter. Ensure PMT gains are reasonable.
* BAKE
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Resuming an acquisition
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