Summary and Setup

This lesson provides an overview of the fundamentals of imaging data that has been acquired through light microscopy experiments. This course is aimed at all researchers who are looking to work with light microscopy imaging data, or to refresh their knowledge on some of the key fundamentals.

Prerequisites

Some prior experience with acquiring/working with light microscopy data would be useful, but isn’t necessary. This course is aimed at anyone who is looking to work with light microscopy imaging data in their research projects.
No python experience is necessary or assumed, but you will get more out of the course if you know some basics e.g. from the online Software Carpentry python course or an in-person course at UCL.
Please bring a laptop (Windows, Mac or Linux) to the lesson. Before the start of the course, please follow the setup instructions so that your laptop is ready to run the material in the lessons.

Download Data

During these lessons we will be using two microscopy images and creating images of our own. Create an empty folder somewhere on your computer to use during the course, this will be your working directory. You can name it anything you like but we recommend microscopy-ideas.

Download 00001_01.ome.tiff (CC BY 4.0 Licence) from the open microscopy environment pages to your working directory.

Download Plate1-Blue-A-12-Scene-3-P3-F2-03.czi (CC BY 4.0 Licence) from the open microscopy environment pages to your working directory.

Software Setup

During these lessons we will use the napari image viewer and the napari-aicsimagio plugin to load imaging data files of various file formats. For beginners we recommend using Anaconda to install and manage these packages. The first step is to download and install Anaconda.

The remaining instructions are written assuming you will be installing and running the software from a terminal. Follow the instructions below to open a terminal on your operating system.

Opening a Terminal

Windows: Click Start > Search for Anaconda Prompt > Click to Open
macOS: Launchpad > Other Application > Terminal
Linux: Open a terminal window

For more information on starting and running Anaconda you may visit the ‘Starting Conda’ section of the Anaconda getting started pages.

Run the following commands in the terminal (lines starting with # are comments and do not need to be run). You can run the commands by copy and pasting them into the terminal and pressing the Enter key.

BASH

# Make sure conda is up to date
conda update -n base conda

# We need to use a python program called pip to install
# the napari-aicsimagio plugin, so we need to configure
# conda to work well with pip
conda config --set pip_interop_enabled True

# This line will create a "virtual environment" (called napari-env)
# that will contain all of the software that will be used in the lessons
conda create -y -n napari-env -c conda-forge python=3.10 bioformats_jar

# Activate the napari-env virtual environment.
# Running this should change the terminal prompt to '(napari-env)'.
conda activate napari-env

# Install napari and plugins using pip
pip install "napari[all]" napari-aicsimageio

Why is there so much text appearing on the screen?

If you’re new to running commands in the terminal you may be alarmed if running some of these commands results in a lot of text output to the screen. This is normal and usually helpful for checking things are working OK. For our purposes you can ignore it. If you see the word ERROR appear near the end (often highlighted in a different colour) it is possible something has gone wrong. In this case the best place to start is any messages in the text expanding on the source of the error.

If you are using an M1/M2 mac, you may encounter errors when running the commands above. If so, try the alternative commands below:

BASH

# Remove the old environment
conda remove --name napari-env --all

# This line will create a "virtual environment" (called napari-env)
# that will contain all of the software that will be used in the lessons
conda create -y -n napari-env -c conda-forge python=3.10 napari pyqt bioformats_jar napari-aicsimageio

# Activate the napari-env virtual environment.
# Running this should change the terminal prompt to '(napari-env)'.
conda activate napari-env

# Install czi file reader
pip install aicspylibczi>=3.0.5 fsspec>=2022.7.1

Post Setup Checks

Before starting the course it is important to check that your setup is working.

Check napari opens a tiff image.

Any napari installation should be able to open a tiff image. Start napari. Note that if you’re already in the napari-env virtual environment from the installation steps it is not necessary to run conda activate napari-env.

BASH

conda activate napari-env
napari

You should now see the napari viewer like this …

A screenshot of the default Napari user interface

Open a file with

File > Open file(s)...

Navigate to the directory where you saved 00001_01.ome.tiff earlier on.
Select 00001_01.ome.tiff and click open.
If you see a Choose reader dialog, select napari builtins.

Expected Output

A screenshot of freshly opened 00001_01.ome.tiff

Close napari: File > exit

Check napari opens a czi image.

During the course we’ll be working with czi images. To open these the napari-aicsimageio plugin is needed. Let’s check the plugin is working.

Start napari. Note that if you’ve already done the previous test (tiff image) then it should not be necessary to run conda activate napari-env. However, it is necessary to restart napari to ensure the image display is correctly formatted.

BASH

conda activate napari-env
napari

File > Open file(s)...

Navigate to the directory where you saved Plate1-Blue-A-12-Scene-3-P3-F2-03.czi to earlier on.
Select Plate1-Blue-A-12-Scene-3-P3-F2-03.czi and click open.
If you see a Choose reader dialog, select napari-aicsimageio.

Expected Output

A screenshot of freshly opened Plate1-Blue-A-12-Scene-3-P3-F2-03.czi

Close napari: File > exit