Image Analysis Using ImageJ

This course will teach the basics of image analysis in ImageJ/Fiji, covering set-up, parameter selection, and output interpretation.

This course is designed to be completed in 1-2 hours. Your progress will be saved and you can return at any time.

Meet your tutor!

Dr Lukasz Chrobok is a lecturer and research fellow at the School of Physiology, Pharmacology & Neuroscience at the University of Bristol. He specialises in circadian rhythm and neurobiology, and has previously delivered webinars on immunofluorescence and immunohistochemistry image analysis.

Contents

Welcome

Welcome to Image Analysis Using ImageJ!

Lecturer and research fellow, Dr. Lukasz Chrobok, will teach you the basics of how to perform image analysis in ImageJ/Fiji, covering set-up, parameter selection, and output interpretation.

Introduction

Introduction to ImageJ

Performing the best quality of image analysis possible starts before you even turn on your computer. This section will talk you through the steps you can take to ensure that the images you analyze will produce robust data consistently.

An introduction to image analysis for immunofluorescence
What is Fiji, and is it just ImageJ?
The importance of good experimental design

How to use Fiji/ImageJ

In this section of the course, we will be learning about the different types of images you can manipulate in ImageJ and we will cover the basics of how to use the software.

Getting started with ImageJ/Fiji
How to add and calibrate a scale bar
Types of images
What is a Z-stack?
Plug-ins
File types
The importance of metadata

Image Manipulation in ImageJ

In this section, we will introduce look-up tables, Z-projections, and show you how to perform intensity and density measurements on your images.

What is the Look-Up Table (LUT)?
How to make composite images and Z-project in Fiji/ImageJ
Measuring intensity and density

Thresholding in ImageJ

Thresholding is a technique that allows image segmentation, allowing users to separate out pixels of interest from background noise.

In this section, we will explain how to perform the different types of thresholding available in ImageJ.

An introduction to thresholding
Global Thresholding
Adaptive Thresholding

How to Normalize Intensity to Background

Performing intensity calculations becomes more complex when working with imperfect sections. Your data can easily be skewed if your background has a high level of intensity.

In this section, we will cover how to factor in the background intensity level when performing intensity calculations to make your data more accurate.

Calculating corrected total cell fluorescence (CTCF)

Using the Cell Counter Plug-in

The source of ImageJ's versatility lies within the wide range of plug-ins available. Many of these plug-ins are incredibly useful, yet still easy to use.

This section will cover how to use the cell counter plug-in for counting analysis of your stained images.

How to use the cell counter plug-in

How to Record Your Own Macros

Macros are automated scripts that allow you to perform a series of tasks without having to select each one individually. They can streamline image processing workflow, saving you time and ensuring consistency in your analyses.

When used appropriately, macros can be powerful tools for auto your workflows. This lesson will focus on how to record your own macros and how to use them.

Macros

Learning Checkpoint

Take this short quiz to recap what we have learned so far.

Summary
Quiz

Congratulations!

Congratulations on completing the Proteintech ECR Training Hub Image Analysis using ImageJ course. You are now ready to start analyzing your images!

Let's recap what we have learned.

Learning Checklist