Protocols. Graphical programming for Icy. a.k.a. programming, for the rest of us
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1 Protocols Graphical programming for Icy a.k.a. programming, for the rest of us
2 Foreword: Reproducible Research Quote: "Results aren't much if they can t be reproduced!" (your boss, your reviewers, your colleagues, you!)
3 Foreword: Reproducible Research Quote: "Results aren't much if they can t be reproduced!" Fact: Most journals now reject papers without proper quantification [...] Image quantification was carefully conducted using Photoshop. [...]
4 Foreword: Reproducible Research Quote: "Results aren't much if they can t be reproduced!" Fact: Most journals now reject papers without proper quantification Conclusion: Image quantification is a protocol in its on right...
5 Foreword: Reproducible Research Quote: "Results aren't much if they can t be reproduced!" Fact: Most journals now reject papers without proper quantification Conclusion: Image quantification is a protocol in its on right... Icy makes these protocols easy to read / write / use / adapt
6 Foreword: Reproducible Research Quote: "Results aren't much if they can t be reproduced!" Fact: Most journals now reject papers without proper quantification Conclusion: Image quantification is a protocol in its on right... Icy makes these protocols easy to read / write / use / adapt Design a protocol once, run on thousands of images Upload your protocol and share with the world (within publications) Download other protocols, run them out-of-the-box Extend any protocol to meet your needs and share/publish again all in just a few clicks, no programming knowledge required.
7 A protocol is a workflow linking processing blocks together Blocks Links
8 Standardised design: all blocks look the same Strong modularity: one block = one task Run protocol until this block (inclusive) and stop Extra actions Block execution priority (starts at 0, click to prioritize) Name (can be modified) Execution state "Drop zones" (link from other blocks) "Drag zones" (link to other blocks) Input parameters (0 or more) Output (0 or more) Arbitrary color code indicates parameter types
9 The protocols editor: New protocol Open a protocol Save / Save as... Embed protocol inside a batch loop Start / Stop the protocol Right click into the workspace to insert a block in the current protocol Search block by name Blocks matching current search. Just drag and drop the block in the workspace to add it. Workspace Documentation of selected block Opened protocol(s) Status bar
10 Blocks are organised by groups all available plug-ins user input (values, images, files, etc.) sequence manipulation operations on regions of interest (read files from / save data to) disk interact with ImageJ (call plugins/macros) create loops (for repetitive tasks) developer tools (more a sandbox really) More ideas on how to tidy things up? Let us know!
11 Question: how would you find the nuclei in this image?
12 Question: how would you find the nuclei in this image? Outline: 1. Extract the channel of interest 2. Clean the data 3. Find an intensity threshold 4. Threshold the image 5. Extract the regions of interest 6. Quantify (notice how generic this outline is )
13 Question: how would you find the nuclei in this image? Outline: 1. Extract the channel of interest 2. Clean the data 3. Find an intensity threshold 4. Threshold the image 5. Extract the regions of interest 6. Quantify Menu: Sequence > Extract Channel NOTE: channel index starts at 0...
14 Question: how would you find the nuclei in this image? Outline: 1. Extract the channel of interest 2. Clean the data 3. Find an intensity threshold 4. Threshold the image 5. Extract the regions of interest 6. Quantify Menu: Blocks > Gaussian Filter Diffuses the intensity contained in each pixel (i.e. makes the image look blurry) Adapt the diffusion to the image noise Too much diffusion: edges fade away!
15 Question: how would you find the nuclei in this image? Outline: 1. Extract the channel of interest 2. Clean the data 3. Find an intensity threshold 4. Threshold the image 5. Extract the regions of interest 6. Quantify Menu: Blocks > KMeans Threshold background foreground Finds the optimal separation(s) between the histogram modes (i.e. intensity classes) 2 modes => 1 threshold (3 => 2, etc.)
16 Question: how would you find the nuclei in this image? Outline: 1. Extract the channel of interest 2. Clean the data 3. Find an intensity threshold 4. Threshold the image 5. Extract the regions of interest 6. Quantify Menu: Blocks > Thresholder Creates a labeled image by classifying pixel intensities according to the threshold(s) 1 threshold => [0;1] (binary) image 2 thresholds => [0;1;2] (labeled) image etc.
17 Question: how would you find the nuclei in this image? Outline: 1. Extract the channel of interest 2. Clean the data 3. Find an intensity threshold 4. Threshold the image 5. Extract the regions of interest 6. Quantify Menu: Blocks > Label Extractor Extracts objects from a labeled image using connected component analysis
18 Question: how would you find the nuclei in this image? Outline: 1. Extract the channel of interest 2. Clean the data 3. Find an intensity threshold 4. Threshold the image 5. Extract the regions of interest 6. Quantify Menu: ROI > ROI Statistics Calculates size, dimensions, intensity statistics, etc.
19 Question: how would you find the nuclei in this image? don t forget me!
20 Question: how would you find the nuclei in this image? click + drag from here
21 Question: how would you find the nuclei in this image? release here
22 Question: how would you find the nuclei in this image? and so forth...
23 Question: how would you find the nuclei in this image? click on the run button
24 Question: how would you find the nuclei in this image? if you want to see something...
25 How about batch processing? all results go in a single file
26 How about batch processing? Results go in a specific file for each image
27 Most plugins have their corresponding Block What if the one you need isn't there (yet)? #1: Leave a comment on the plug-in's page online Leave a comment & rating
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