// Elastin Fiber Length Quantification. 

// Epi vs. enothelium maybe? 

// Either way, a quick search of the Image.sc forums pointed towards the 
//	Ridge Detection plugin. Looks like it's going to work fine. 
// https://imagej.net/Ridge_Detection

// ===============================
// Written by Wilson R Adams | Vanderbilt Biophotonics Center | 16 Feb 2021
// ==============================

input = getTitle(); 
elastinlengths(input);

function elastinlengths(img) { 
// Uses ridge detection to segment out elastin fiber features and quantifies their lengths.
	// Prep
	roiManager("reset");

	imdir = getInfo("image.directory");
	pardir = File.getParent(imdir);
	resdir = imdir+File.separator+"Results";
	File.makeDirectory(resdir);	// make results directory
	selectWindow(img);
	name = File.nameWithoutExtension; // get filename variable to rename outputs

	// Start Processing - Prep image for function
	run("Duplicate...", " ");
	wrk = getTitle();
	run("Enhance Contrast", "saturated=0.35");
	run("Enhance Contrast", "saturated=0.35");
	//run("Gaussian Blur...", "sigma=1");	// does this help? - no not really. 
	run("8-bit"); // Needs 8bit grayscale to run plugin

	// Run function to get fiber length
	run("Ridge Detection", "line_width=3 high_contrast=100 low_contrast=0 extend_line displayresults add_to_manager make_binary method_for_overlap_resolution=NONE sigma=1.37 lower_threshold=0 upper_threshold=4.25 minimum_line_length=10 maximum=0"); // Rejecting fibers shorter than 10px

	close("Junctions");
	close("Results");
	selectWindow("Summary");
	saveAs("Results", resdir+File.separator+name+"_Summary.csv");
	close(name+"_Summary.csv");
	roiManager("save", resdir+File.separator+name+"_ROI.zip");
	selectWindow(wrk+" Detected segments");
	save(resdir+File.separator+wrk+" Detected segments.tif");
	close();
	close();
	roiManager("reset");
	close();
};