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The Cell Proliferation recipe in Aivia detects cell regions and measures the change in confluency over time. The recipe applies a machine learning-based algorithm for cell segmentation and works for both phase contrast and fluorescence microscopy images.



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Parameters and Presets

Parameters

Recipe parameters for Cell Proliferation and their descriptions are summarized in the table below.

Preset GroupParameter NameMin ValueMax ValueDescription
Detection


Background Flattening Factor

(Enhance Image only)

0100

Adjusts the sensitivity of the background removal operation; a lower value will preserve larger objects and more background variations

Contrast Threshold (Enhance Image)

Intensity Threshold (Skip Enhance Image)

0

255 (8-bit)

65,535 (16-bit)

Adjusts the detection sensitivity on the segmentation image;

  • When Enhance Image is selected, the threshold will be applied to the enhanced image with the maximum value set to 255 regardless of image bit-depth
  • When Skip Enhance Image is selected, the threshold will be applied to the input image

A lower value will detect bigger and more objects

Subset Filtering


Fill Holes Size01,000,000Adjusts the maximum size threshold for filling in gaps inside a detected object; a lower value will preserve more holes in the detection
Smoothing Factor0100Adjusts the amount of smoothing applied to the outline of the detected objects; a lower value will preserve more of the object's morphological features
Min Object Size01,000,000Specifies the minimum size of objects to be included in the analysis results based on the area of detected regions



Presets

There are two preset groups in the recipe: Detection and Subset Filtering; each group has three pre-configured parameter groupings to help you get started on the analysis. The default preset values are as follows:


Detection

Parameter NameLowMediumHigh
Background Flattening Factor506063

Contrast Threshold

(Enhance Image only)

20

20

20

Intensity Threshold

(Skip Enhance Image only)

51 (8-bit)

13,107 (16-bit)

20 (8-bit)

5,243 (16-bit)

5 (8-bit)

1,311 (16-bit)




Subset Filtering

Parameter NameSmallMediumLarge
Fill Holes Size505001,000
Smoothing Factor124
Object Size505005,000




Measurements

The Cell Proliferation recipe generates morphological measurements for the confluency of the detected regions. You can add additional measurements to the analysis results by using the Measurement Tool in Aivia. The measurements generated by the recipe are as follows:


MorphologicalSummary
  • Area
  • Percent Total Image
  • Cell region area over time
  • Percent area of cells over time




Tutorial

Before beginning the tutorial, please download the Cell Proliferation Demo image. For information on how to select presets or modify parameter values, please refer to the tutorial on how to use the Recipe Console.

  1. Unzip the demo file and load the demo image, CellProDemo.tif, into Aivia
  2. In the Recipe Console, click on the Recipe selection dropdown menu and select the Cell Proliferation recipe
  3. Select the High preset for the Detection group and the Medium preset for the Subset Filtering group
  4. Click on the caret Image Modified to the left of the Subset Filtering preset group to show the preset parameters
  5. Change the Subset Filtering preset parameter below as follows, leaving the others intact:
    • Fill Holes Size: 500
  6. Click the Start button or press the F4 key on your keyboard to begin applying the recipe to the image


The detected cell region outline will be overlaid on the image.


Results

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Cell Proliferation results




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