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Detecting neurites

Introduction

The guide explains you to detect neurons structures using the tracing module. This module offers algorithms to automatically segment neurons structures based on signal backbone seeds (local maxima) detection and their linking in a logical tree-like architecture. From the segments, several features are afterward measured.

Workflow

Demo dataset

The data set is a multi-dimensional, discrete representation of your real sample volume. It can be structured as a z-series of planes (optical slices) of multiple channels (dyes) in a temporal sequence of time points located at multiple spatial positions. Usually, the dataset shows a single experimental situation. A complete experiment can be composed by several datasets. The datasets are available as graphic files saved in plenty of file formats (standard formats as well as proprietary formats).

The link for the specific demo dataset for this guide is displayed below. All datasets are listed here: https://demodata.arivis.com

Downloading the demo dataset

  1. To download the demo dataset, click http://demodata.arivis.com/data/arivisVision4D-DemoData-SamplePipelines-Trace.zip.
  2. The download is starting automatically.
  3. Create a new folder on your local disk. Move the *.zip file from the download folder inside it.
  4. Unzip the folder.

Opening the demo dataset

  1. Click File > Open...
  2. Select the *.sis file from the Windows Explorer.
  3. The demo dataset is displayed in the viewer. The dataset is visualized according to the current rendering setting parameters.

Activating the Sample Pipeline

  1. In the Shortcut Toolbar, click Analysis Panel .
  2. In the Sample Pipelines list, double-click the Detect Neurites Pipeline.
  3. If you have activated a pipeline, it will be replaced by the new one.

You can open the appropriate How to guide. By hovering over a pipeline, this button is displayed. When clicking on it, the option Open How to appears.

Pipeline operations layout

The following operations are part of the pipeline.

Input ROI

This operation allows to select the region of interest (ROI). ROI defines the dataset subarea that will be processed and analyzed by the pipeline.

Neurite Tracer

Automatic tubular-like structures detection based on the threshold-based reconstruction algorithm.

Store Objects

Store the detected segments (tag) in the active dataset.

Executing the pipeline

You can execute the pipeline step by step or in a single run. To do this, use the executing buttons in the Pipeline toolbar.

Executing Buttons in the Pipeline toolbar
Executing Buttons in the Pipeline toolbar

Executing step by step

You can execute the pipeline step by step (back and forth). This method allows to run and undo a single operation. You can either use the executing buttons in the Pipeline toolbar or in the Operation toolbar to go through the operation list.

  1. To run the single operation, click .
  2. To undo the single operation, click .
    Note: Undo the last operation executed if you need to change the operation settings.

Executing in a single run

As alternative to executing step by step, execute the pipeline in a single run.

  1. To run the whole pipeline, click .
  2. To stop the pipeline execution, click .

Operation status

Executing buttons in the Operation toolbar after executing
Executing buttons in the Operation toolbar after executing

When the operation is running, this icon is shown.

When the operation is completed, this icon is shown.

Viewing the results

  1. If not already visible, open the Objects dialog. In Shortcut Toolbar, click Objects table .
  2. Measurements are now visible in the objects table.
  3. To color the results more meaningfully, click Set Object Style.
  4. Click Apply to: Traces.
  5. To select the trace color options, select one item from the Color by list.

    For some visualization examples, see Visualization examples.
  6. To set further style settings, set the parameters in the Visualization section.

    For some visualization examples, see Visualization examples.

Refer to the Help (F1) for more Information.

Visualization examples

Color by structure - Cone

Color by structure - Approximation

Color by depth - Approximation

Color by depth - Skeleton

Color by depth - Cone

Modifying the current pipeline

You can modify the pipeline to adapt to another datasets. Therefore, all the pipeline parameters should be set according to the new dataset features.

Previewing the results

For all operations the preview is available in 2D, for some also in 3D.

  1. Switch from 2D Viewer to 4D Viewer in the Viewer Type Switch.
  2. To preview the operation results, click Preview in the Operation toolbar.

Use the Navigator panel in the Panel Sidebar to select the preview z plane and/or time points.

Adjusting the operations

The parameters of each operation are described here: Pipeline operations layout

  1. You have opened the pipeline.
  1. Go to the operation you want to change.
  2. Set the appropriate parameters as desired.
  3. Execute the pipeline (see Executing the pipeline).

Adding or removing operations

You can add or remove operations from the sample pipeline.

  1. Click + Add operation...
  2. The operation list is grouped in four groups by their typology.
  3. To add the operation to the current pipeline, double-click it. Alternatively, drag & drop the operation to the current Pipeline.
    Note:
    The operation cannot be added during the Pipeline execution.
  4. The operation will be inserted at the end of the group of operations to which it belongs.
    Voxel operations are positioned before the segment generation. Store operations are always put at the end of the Pipeline.
  5. To remove an operation, click Close at the Operation toolbar.

Editing the traces

The tubular structures segmented (traces) by the operation can be edited. Branches, terminal points, branches points, and other trace components are accessible for modification, optimizations, and improvements.

  1. In the Shortcut toolbar, click Trace Tool .
  2. An Option Bar below the Shortcut Toolbar is displayed. Here you can find various tools with which you can draw or edit traces.

  3. Most of these tools can be activated using simple shortcuts to facilitate the workflow.

Refer to the Help (F1) for more details.

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