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CellProfiler Pipeline: http://www.cellprofiler.org
Version:5
DateRevision:400
GitHash:
ModuleCount:14
HasImagePlaneDetails:False
Images:[module_num:1|svn_version:'Unknown'|variable_revision_number:2|show_window:False|notes:['To begin creating your project, use the Images module to compile a list of files and/or folders that you want to analyze. You can also specify a set of rules to include only the desired files in your selected folders.']|batch_state:array([], dtype=uint8)|enabled:True|wants_pause:False]
:
Filter images?:Images only
Select the rule criteria:and (extension does isimage) (directory doesnot containregexp "[\\\\\\\\/]\\\\.")
Metadata:[module_num:2|svn_version:'Unknown'|variable_revision_number:6|show_window:False|notes:['The Metadata module optionally allows you to extract information describing your images (i.e, metadata) which will be stored along with your measurements. This information can be contained in the file name and/or location, or in an external file.']|batch_state:array([], dtype=uint8)|enabled:True|wants_pause:False]
Extract metadata?:No
Metadata data type:Text
Metadata types:{}
Extraction method count:1
Metadata extraction method:Extract from file/folder names
Metadata source:File name
Regular expression to extract from file name:^(?P<Plate>.*)_(?P<Well>[A-P][0-9]{2})_s(?P<Site>[0-9])_w(?P<ChannelNumber>[0-9])
Regular expression to extract from folder name:(?P<Date>[0-9]{4}_[0-9]{2}_[0-9]{2})$
Extract metadata from:All images
Select the filtering criteria:and (file does contain "")
Metadata file location:Elsewhere...|
Match file and image metadata:[]
Use case insensitive matching?:No
Metadata file name:
Does cached metadata exist?:No
NamesAndTypes:[module_num:3|svn_version:'Unknown'|variable_revision_number:8|show_window:False|notes:['DNA: DNA stained with DAPI', 'PH3: An antibody for phosphorylated histone H3 correlated with mitosis', 'cellbody: ']|batch_state:array([], dtype=uint8)|enabled:True|wants_pause:False]
Assign a name to:Images matching rules
Select the image type:Grayscale image
Name to assign these images:DNA
Match metadata:[]
Image set matching method:Order
Set intensity range from:Image metadata
Assignments count:3
Single images count:0
Maximum intensity:255.0
Process as 3D?:No
Relative pixel spacing in X:1.0
Relative pixel spacing in Y:1.0
Relative pixel spacing in Z:1.0
Select the rule criteria:and (file does contain "d0.tif")
Name to assign these images:DNA
Name to assign these objects:Cell
Select the image type:Grayscale image
Set intensity range from:Image metadata
Maximum intensity:255.0
Select the rule criteria:and (file does contain "d1.tif")
Name to assign these images:PH3
Name to assign these objects:Cell
Select the image type:Grayscale image
Set intensity range from:Image metadata
Maximum intensity:255.0
Select the rule criteria:and (file does contain "d2.tif")
Name to assign these images:cellbody
Name to assign these objects:Cell
Select the image type:Grayscale image
Set intensity range from:Image metadata
Maximum intensity:255.0
Groups:[module_num:4|svn_version:'Unknown'|variable_revision_number:2|show_window:False|notes:['The Groups module optionally allows you to split your list of images into image subsets (groups) which will be processed independently of each other. Examples of groupings include screening batches, microtiter plates, time-lapse movies, etc.']|batch_state:array([], dtype=uint8)|enabled:True|wants_pause:False]
Do you want to group your images?:No
grouping metadata count:1
Metadata category:None
IdentifyPrimaryObjects:[module_num:5|svn_version:'Unknown'|variable_revision_number:14|show_window:True|notes:[]|batch_state:array([], dtype=uint8)|enabled:True|wants_pause:False]
Select the input image:DNA
Name the primary objects to be identified:Nuclei
Typical diameter of objects, in pixel units (Min,Max):8,80
Discard objects outside the diameter range?:Yes
Discard objects touching the border of the image?:Yes
Method to distinguish clumped objects:Intensity
Method to draw dividing lines between clumped objects:Intensity
Size of smoothing filter:10
Suppress local maxima that are closer than this minimum allowed distance:7.0
Speed up by using lower-resolution image to find local maxima?:Yes
Fill holes in identified objects?:After declumping only
Automatically calculate size of smoothing filter for declumping?:Yes
Automatically calculate minimum allowed distance between local maxima?:Yes
Handling of objects if excessive number of objects identified:Continue
Maximum number of objects:500
Display accepted local maxima?:No
Select maxima color:Blue
Use advanced settings?:No
Threshold setting version:11
Threshold strategy:Global
Thresholding method:Minimum Cross-Entropy
Threshold smoothing scale:1.3488
Threshold correction factor:1.0
Lower and upper bounds on threshold:0.0,1.0
Manual threshold:0.0
Select the measurement to threshold with:None
Two-class or three-class thresholding?:Two classes
Assign pixels in the middle intensity class to the foreground or the background?:Foreground
Size of adaptive window:50
Lower outlier fraction:0.05
Upper outlier fraction:0.05
Averaging method:Mean
Variance method:Standard deviation
# of deviations:2.0
Thresholding method:Otsu
IdentifyPrimaryObjects:[module_num:6|svn_version:'Unknown'|variable_revision_number:14|show_window:True|notes:[]|batch_state:array([], dtype=uint8)|enabled:True|wants_pause:False]
Select the input image:PH3
Name the primary objects to be identified:PH3
Typical diameter of objects, in pixel units (Min,Max):7,80
Discard objects outside the diameter range?:Yes
Discard objects touching the border of the image?:Yes
Method to distinguish clumped objects:Intensity
Method to draw dividing lines between clumped objects:Intensity
Size of smoothing filter:10
Suppress local maxima that are closer than this minimum allowed distance:7.0
Speed up by using lower-resolution image to find local maxima?:Yes
Fill holes in identified objects?:After declumping only
Automatically calculate size of smoothing filter for declumping?:Yes
Automatically calculate minimum allowed distance between local maxima?:Yes
Handling of objects if excessive number of objects identified:Continue
Maximum number of objects:500
Display accepted local maxima?:No
Select maxima color:Blue
Use advanced settings?:Yes
Threshold setting version:11
Threshold strategy:Global
Thresholding method:Otsu
Threshold smoothing scale:1.3488
Threshold correction factor:1.0
Lower and upper bounds on threshold:0.0,1.0
Manual threshold:0.0
Select the measurement to threshold with:None
Two-class or three-class thresholding?:Three classes
Assign pixels in the middle intensity class to the foreground or the background?:Foreground
Size of adaptive window:50
Lower outlier fraction:0.05
Upper outlier fraction:0.05
Averaging method:Mean
Variance method:Standard deviation
# of deviations:2.0
Thresholding method:Otsu
RelateObjects:[module_num:7|svn_version:'Unknown'|variable_revision_number:5|show_window:True|notes:[]|batch_state:array([], dtype=uint8)|enabled:True|wants_pause:False]
Parent objects:Nuclei
Child objects:PH3
Calculate child-parent distances?:None
Calculate per-parent means for all child measurements?:No
Calculate distances to other parents?:No
Do you want to save the children with parents as a new object set?:No
Name the output object:None
Parent name:None
Parent name:None
IdentifySecondaryObjects:[module_num:8|svn_version:'Unknown'|variable_revision_number:10|show_window:True|notes:[]|batch_state:array([], dtype=uint8)|enabled:True|wants_pause:False]
Select the input objects:Nuclei
Name the objects to be identified:Cells
Select the method to identify the secondary objects:Propagation
Select the input image:cellbody
Number of pixels by which to expand the primary objects:10
Regularization factor:0.05
Discard secondary objects touching the border of the image?:No
Discard the associated primary objects?:No
Name the new primary objects:FilteredNuclei
Fill holes in identified objects?:Yes
Threshold setting version:11
Threshold strategy:Global
Thresholding method:Minimum Cross-Entropy
Threshold smoothing scale:0.0
Threshold correction factor:0.8
Lower and upper bounds on threshold:0.0,1.0
Manual threshold:0.0
Select the measurement to threshold with:None
Two-class or three-class thresholding?:Three classes
Assign pixels in the middle intensity class to the foreground or the background?:Foreground
Size of adaptive window:50
Lower outlier fraction:0.05
Upper outlier fraction:0.05
Averaging method:Mean
Variance method:Standard deviation
# of deviations:2.0
Thresholding method:Otsu
IdentifyTertiaryObjects:[module_num:9|svn_version:'Unknown'|variable_revision_number:3|show_window:True|notes:[]|batch_state:array([], dtype=uint8)|enabled:True|wants_pause:False]
Select the larger identified objects:Cells
Select the smaller identified objects:Nuclei
Name the tertiary objects to be identified:Cytoplasm
Shrink smaller object prior to subtraction?:Yes
MeasureObjectIntensity:[module_num:10|svn_version:'Unknown'|variable_revision_number:4|show_window:True|notes:[]|batch_state:array([], dtype=uint8)|enabled:True|wants_pause:False]
Select images to measure:DNA, PH3
Select objects to measure:Nuclei, Cells, Cytoplasm
MeasureObjectSizeShape:[module_num:11|svn_version:'Unknown'|variable_revision_number:3|show_window:True|notes:[]|batch_state:array([], dtype=uint8)|enabled:True|wants_pause:False]
Select object sets to measure:Nuclei, Cells, Cytoplasm
Calculate the Zernike features?:Yes
Calculate the advanced features?:No
OverlayOutlines:[module_num:12|svn_version:'Unknown'|variable_revision_number:4|show_window:True|notes:[]|batch_state:array([], dtype=uint8)|enabled:True|wants_pause:False]
Display outlines on a blank image?:No
Select image on which to display outlines:DNA
Name the output image:OrigOverlay
Outline display mode:Color
Select method to determine brightness of outlines:Max of image
How to outline:Thick
Select outline color:#0080FF
Select objects to display:Cells
Select outline color:blue
Select objects to display:Nuclei
Select outline color:yellow
Select objects to display:PH3
SaveImages:[module_num:13|svn_version:'Unknown'|variable_revision_number:15|show_window:True|notes:[]|batch_state:array([], dtype=uint8)|enabled:True|wants_pause:False]
Select the type of image to save:Image
Select the image to save:OrigOverlay
Select method for constructing file names:From image filename
Select image name for file prefix:DNA
Enter single file name:OrigBlue
Number of digits:4
Append a suffix to the image file name?:Yes
Text to append to the image name:_Overlay
Saved file format:png
Output file location:Default Output Folder|
Image bit depth:8-bit integer
Overwrite existing files without warning?:Yes
When to save:Every cycle
Record the file and path information to the saved image?:Yes
Create subfolders in the output folder?:No
Base image folder:Elsewhere...|
How to save the series:T (Time)
ExportToSpreadsheet:[module_num:14|svn_version:'Unknown'|variable_revision_number:13|show_window:True|notes:[]|batch_state:array([], dtype=uint8)|enabled:True|wants_pause:False]
Select the column delimiter:Comma (",")
Add image metadata columns to your object data file?:No
Add image file and folder names to your object data file?:No
Select the measurements to export:No
Calculate the per-image mean values for object measurements?:No
Calculate the per-image median values for object measurements?:No
Calculate the per-image standard deviation values for object measurements?:No
Output file location:Default Output Folder|
Create a GenePattern GCT file?:No
Select source of sample row name:Metadata
Select the image to use as the identifier:None
Select the metadata to use as the identifier:None
Export all measurement types?:Yes
Press button to select measurements:
Representation of Nan/Inf:NaN
Add a prefix to file names?:No
Filename prefix:MyExpt_
Overwrite existing files without warning?:Yes
Data to export:Do not use
Combine these object measurements with those of the previous object?:No
File name:DATA.csv
Use the object name for the file name?:Yes