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Course 4 – Advanced Image Analysis & Processing April 2015 1

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Page 1: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Course 4 –Advanced Image Analysis & Processing

April 2015

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Page 2: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

From Survey Results

• Heterogenous audience

• Day 1 : ImageJ -Macro : language, batch, use of ActionBar.

• Day2 : ImageJ - Plugins : install, writing, use of Eclipse.

• Day 3 : Matlab & CellProfiler, how to use them for image analysis, comparison to ImageJ

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Page 3: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Fiji

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Page 4: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

FijiPlugins

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http://fiji.sc/Fiji

Page 5: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Get Action Bar Plugin

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Page 6: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Add Action Bar Plugin

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Fiji.app\plugins

Paste :

Page 7: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Add Action Bar Plugin

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Fiji.app\plugins

Paste :

Page 8: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

My first Action Bar

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Page 9: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

My first Action Bar

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Fiji.app\plugins\ActionBar

Page 10: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

My first Action Bar

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Refresh Menus

Page 11: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

My first Action Bar

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Fiji.app\plugins\ActionBar

Page 12: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

My first Action Bar

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Create

Page 13: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

My first Action Bar

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Paste

Page 14: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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ChangeFile

Format

Colored texteasier to debug

Page 15: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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Not Saved

Page 16: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Save Your Action Bar

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My_Action_Bar.ijm

Page 17: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Action Barbasic functions

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Page 18: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Action Barbasic functions

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Page 19: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Action Barchange Icon

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New icon file name

Page 20: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Action Barchange Icon

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Page 21: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Action Barchange Label

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label

Page 22: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Action Barchange Label

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label

Page 23: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Action when clic on button

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Clic

Actionto perform

//using macro functions

Page 24: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Macro Functions

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Recorder Website

OR

Page 25: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Macro Functions

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OR

Recorder Website

Page 26: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Record Functions

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Page 27: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Record Functions

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RGB

Macro language

Page 28: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Record Functions

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Actionto perform

//using macro functions

// start of : line

Page 29: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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of the button lineendstart

Page 30: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Use a macro : on an image

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DAPI

H2B-RFPEdU-488

Create a nuclei mask from DAPI

Get for each nuclei :- the «Area» and - the «Intensitiy» of the Red and Green Channel

Create a macro to analyse Red and Green staining.

You have 30min!

Page 31: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

How to write a macro

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Template: tiny.cc\adip-1stStepMacro_blank

Page 32: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

How to write a macro

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Solution : tiny.cc\adip-1stStepMacro

Page 33: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Use a macro : on a folder

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Page 34: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Use a macro : on a folder

Process>Batch>Macro

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Page 35: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Use a macro on a folder

Process>Batch>Macro

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Necessary to handle the processing of several images

Solution : tiny.cc\adip-2ndStepMacro

Page 36: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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Use a macro on a folder

Process>Batch>Macro + GUI for parameters

Page 37: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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Use a macro on a folder

Process>Batch>Macro + GUI for parameters

• How to ask user to set/select parameters

Starts with : Dialog.create(); Website

Page 38: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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Use a macro on a folder

Process>Batch>Macro + GUI for parameters

Solution : tiny.cc\adip-3rdStepMacro

Page 39: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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Use a macro on a folder

Process>Batch>Macro + GUI for parameters

Page 40: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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Use a macro on a folder

Process>Batch>Macro + GUI for parameters

How to specify the parameters once and use them for all images ?

Page 41: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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• Keep the same Macro• Modify it so it can process every file in a folder!

• You will need:– The path of the folder containing images.– The list of the files.

• You will have to :– Create a new folder to save images (mask)– Process every file.– Save the created mask of every image.

Use a macro on a folder

Macro: process image of a folder

Page 42: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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• Keep the same Macro• Modify it so it can process every file in a folder!

• You will need:– The path of the folder containing images. getDirectory– The list of the files. getFileList

• You will have to :– Create a new folder to save images (mask) …– Process every file …– Save the created mask of every image …

Use a macro on a folder

Macro: process image of a folder

Page 43: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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Use a macro on a folder

Macro: process image of a folder

Page 44: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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What happens if : there is non-image File within the folder?

To prevent issue, we’ll use a functionto control that files are images.

Use a macro on a folder

Macro: process image of a folder

Solution : tiny.cc\adip-4thStepMacro

Page 45: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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Use a macro in a Action Bar!

Macro: process image of a folder

//using macro functions

We can use our own <macro> code

Page 46: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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Use a macro in a Action Bar!

on 1 image

adip-1stStepMacro.ijmWe can use our own <macro> code

Solution : tiny.cc\MyImproved_action_bar_step1.ijm

Page 47: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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Use a macro in a Action Bar!

on 1 folder

adip-4thStepMacro.ijmWe can use our own <macro> code

Solution : tiny.cc\MyImproved_action_bar_step2.ijm

Page 48: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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Use a macro in a Action Bar!

on 1 folder

adip-4thStepMacro.ijmWe can use our own <macro> code

This function can be necessaryfor other buttons of the Action Bar

Page 49: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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Use a macro in a Action Bar!

codelibrary

contains functions that every <button> can access<codeLibrary>

Which other functions can we add to the <codeLibrary> ?

Solution : tiny.cc\MyImproved_action_bar_step3.ijm

Page 50: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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Use a macro in a Action Bar!

codelibrary

the common part when processing1 image and 1 folder

Page 51: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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Use a macro in a Action Bar!

codelibrary

Page 52: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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Solution : tiny.cc\MyImproved_action_bar_step4.ijm

Use a macro in a Action Bar!

codelibrary

Page 53: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

• We need to:– Store data (text, numbers…)– Retrieve these data

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Use a macro in a Action Bar!

Parameters Windows : function

Page 54: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

Use a macro in a Action Bar!

Parameters Windows : function

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Container will be an ImageJ text window

Page 55: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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Use a macro in a Action Bar!

Parameters Windows : function

Page 56: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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Use a macro in a Action Bar!

Parameters Windows : function

Page 57: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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Set them first !

Retrieve data

Use a macro in a Action Bar!

Parameters Windows : modify action bar

Solution : tiny.cc\MyImproved_action_bar_step5-0.ijm

Page 58: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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Use a macro in a Action Bar!

Parameters Windows : setData

Solution : tiny.cc\MyImproved_action_bar_step5-1.ijm

Set Parameters <button>

Page 59: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

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Use a macro in a Action Bar!

Parameters Windows : setData

Solution : tiny.cc\MyImproved_action_bar_step5.ijm

Use it for the Folder <button>

Page 60: Course 4 Advanced Image Analysis & Processing · 2018-04-12 · Advanced Image Analysis & Processing April 2015 1. From Survey Results •Heterogenous audience •Day 1 : ImageJ -Macro

That’s all Folks!

Tomorrow : JaaavvaaaAAAArrrgggghhh

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