Creating Histograms#

Questions

  • How can we create grayscale and colour histograms to understand the distribution of colour values in an image?

Objectives

  • Explain what a histogram is.

  • Load an image in grayscale format.

  • Create and display grayscale and colour histograms for entire images.

  • Create and display grayscale and colour histograms for certain areas of images, via masks.

In this episode, we will learn how to use scikit-image functions to create and display histograms for images.

First, import the packages needed for this episode

import imageio.v3 as iio
import ipympl
import matplotlib.pyplot as plt
import numpy as np
import skimage as ski

%matplotlib widget

Introduction to Histograms#

As it pertains to images, a histogram is a graphical representation showing how frequently various colour values occur in the image. We saw in the Image Basics episode that we could use a histogram to visualise the differences in uncompressed and compressed image formats. If your project involves detecting colour changes between images, histograms will prove to be very useful, and histograms are also quite handy as a preparatory step before performing thresholding.

Grayscale Histograms#

We will start with grayscale images, and then move on to colour images. We will use this image of a plant seedling as an example:

Plant seedling

Here we load the image in grayscale instead of full colour, and display it:

# read the image of a plant seedling as grayscale from the outset
plant_seedling = iio.imread(uri="data/plant-seedling.jpg", mode="L")

# convert the image to float dtype with a value range from 0 to 1
plant_seedling = ski.util.img_as_float(plant_seedling)

# display the image
fig, ax = plt.subplots()
ax.imshow(plant_seedling, cmap="gray")
<matplotlib.image.AxesImage at 0x7fe104979e90>

Again, we use the iio.imread() function to load our image. Then, we convert the grayscale image of integer dtype, with 0-255 range, into a floating-point one with 0-1 range, by calling the function ski.util.img_as_float. We can also calculate histograms for 8 bit images as we will see in the subsequent exercises.

We now use the function np.histogram to compute the histogram of our image which, after all, is a NumPy array:

# create the histogram
histogram, bin_edges = np.histogram(plant_seedling, bins=256, range=(0, 1))

The parameter bins determines the number of “bins” to use for the histogram. We pass in 256 because we want to see the pixel count for each of the 256 possible values in the grayscale image.

The parameter range is the range of values each of the pixels in the image can have. Here, we pass 0 and 1, which is the value range of our input image after conversion to floating-point.

The first output of the np.histogram function is a one-dimensional NumPy array, with 256 rows and one column, representing the number of pixels with the intensity value corresponding to the index. I.e., the first number in the array is the number of pixels found with intensity value 0, and the final number in the array is the number of pixels found with intensity value 255. The second output of np.histogram is an array with the bin edges and one column and 257 rows (one more than the histogram itself). There are no gaps between the bins, which means that the end of the first bin, is the start of the second and so on. For the last bin, the array also has to contain the stop, so it has one more element, than the histogram.

Next, we turn our attention to displaying the histogram, by taking advantage of the plotting facilities of the Matplotlib library.

# configure and draw the histogram figure
fig, ax = plt.subplots()
ax.set_title("Grayscale Histogram")
ax.set_xlabel("grayscale value")
ax.set_ylabel("pixel count")
ax.set_xlim([0.0, 1.0])  # <- named arguments do not work here

ax.plot(bin_edges[0:-1], histogram)  # <- or here
[<matplotlib.lines.Line2D at 0x7fe104797910>]

We create the plot with plt.subplots(), then label the figure and the coordinate axes with ax.set_title(), ax.set_xlabel(), and ax.set_ylabel() functions. The last step in the preparation of the figure is to set the limits on the values on the x-axis with the ax.set_xlim([0.0, 1.0]) function call.

Variable-length argument lists

Note that we cannot used named parameters for the ax.set_xlim() or ax.plot() functions. This is because these functions are defined to take an arbitrary number of unnamed arguments. The designers wrote the functions this way because they are very versatile, and creating named parameters for all of the possible ways to use them would be complicated.

Finally, we create the histogram plot itself with ax.plot(bin_edges[0:-1], histogram). We use the left bin edges as x-positions for the histogram values by indexing the bin_edges array to ignore the last value (the right edge of the last bin).

Histograms in Matplotlib

Matplotlib provides a dedicated function to compute and display histograms: ax.hist(). We will not use it in this lesson in order to understand how to calculate histograms in more detail. In practice, it is a good idea to use this function, because it visualises histograms more appropriately than ax.plot(). Here, you could use it by calling ax.hist(image.flatten(), bins=256, range=(0, 1)) instead of np.histogram() and ax.plot() (*.flatten() is a NumPy function that converts our two-dimensional image into a one-dimensional array).

Exercise: Using a mask for a histogram (15 min)#

Looking at the histogram above, you will notice that there is a large number of very dark pixels, as indicated in the chart by the spike around the grayscale value 0.12. That is not so surprising, since the original image is mostly black background. What if we want to focus more closely on the leaf of the seedling? That is where a mask enters the picture!

First, hover over the plant seedling image with your mouse to determine the (x, y) coordinates of a bounding box around the leaf of the seedling. Then, using techniques from the Drawing and Bitwise Operations episode, create a mask with a white rectangle covering that bounding box.

After you have created the mask, apply it to the input image before passing it to the np.histogram function.

Colour Histograms#

We can also create histograms for full colour images, in addition to grayscale histograms. We have seen colour histograms before, in the Image Basics episode. A program to create colour histograms starts in a familiar way:

# read original image, in full color
plant_seedling = iio.imread(uri="data/plant-seedling.jpg")

# display the image
fig, ax = plt.subplots()
ax.imshow(plant_seedling)
<matplotlib.image.AxesImage at 0x7fe104bec8d0>

We read the original image, now in full colour, and display it. Next, we create the histogram. Since we have a colour image, we use np.histogram three times, once for each of the channels (red, green, blue). We use a for loop to do this, which is a common Python programming pattern. To produce the colours in the histogram plot, we use a tuple variable called colors:

colors = ("red", "green", "blue")

This specifies that the line for the red channel should be coloured red, the green channel line should be green, and the blue channel line should be blue.

# tuple to select colors of each channel line
colors = ("red", "green", "blue")

# create the histogram plot, with three lines, one for each color
fig, ax = plt.subplots()
ax.set_xlim([0, 256])
for channel_id, color in enumerate(colors):
    histogram, bin_edges = np.histogram(
        plant_seedling[:, :, channel_id], bins=256, range=(0, 256)
    )
    ax.plot(bin_edges[0:-1], histogram, color=color)

ax.set_title("Color Histogram")
ax.set_xlabel("Color value")
ax.set_ylabel("Pixel count")
Text(0, 0.5, 'Pixel count')

We will draw the histogram line for each channel in a different colour, and so we create a tuple of the colours to use for the three lines with the

colors = ("red", "green", "blue")

line of code. Then, we limit the range of the x-axis with the ax.set_xlim() function call.

Next, we use the for control structure to iterate through the three channels, plotting an appropriately-coloured histogram line for each. This may be new Python syntax for you, so we will take a moment to discuss what is happening in the for statement.

The Python built-in enumerate() function takes a list and returns an iterator of tuples, where the first element of the tuple is the index and the second element is the element of the list.

Iterators, tuples, and enumerate()

In Python, an iterator, or an iterable object, is something that can be iterated over with the for control structure. A tuple is a sequence of objects, just like a list. However, a tuple cannot be changed, and a tuple is indicated by parentheses instead of square brackets. The enumerate() function takes an iterable object, and returns an iterator of tuples consisting of the 0-based index and the corresponding object.

For example, consider this small Python program:

list = ("a", "b", "c", "d", "e")

for x in enumerate(list):
    print(x)

Executing this program would produce the following output:

(0, 'a')
(1, 'b')
(2, 'c')
(3, 'd')
(4, 'e')

In our colour histogram program, we are using a tuple, (channel_id, color), as the for variable. The first time through the loop, the channel_id variable takes the value 0, referring to the position of the red colour channel, and the color variable contains the string "red". The second time through the loop the values are the green channels index 1 and "green", and the third time they are the blue channel index 2 and "blue".

Inside the for loop, our code looks much like it did for the grayscale example. We calculate the histogram for the current channel with the

histogram, bin_edges = np.histogram(image[:, :, channel_id], bins=256, range=(0, 256))

function call, and then add a histogram line of the correct colour to the plot with the

ax.plot(bin_edges[0:-1], histogram, color=color)

function call. Note the use of our loop variables, channel_id and color.

Finally we label our axes and display the histogram, shown here:

Colour histogram

Exercise: Colour histogram with a mask (25 min)#

We can also apply a mask to the images we apply the colour histogram process to, in the same way we did for grayscale histograms. Consider this image of a well plate, where various chemical sensors have been applied to water and various concentrations of hydrochloric acid and sodium hydroxide:

# read the image
wellplate = iio.imread(uri="data/wellplate-02.tif")

# display the image
fig, ax = plt.subplots()
ax.imshow(wellplate)

Well plate image

Suppose we are interested in the colour histogram of one of the sensors in the well plate image, specifically, the seventh well from the left in the topmost row, which shows Erythrosin B reacting with water.

Hover over the image with your mouse to find the centre of that well and the radius (in pixels) of the well. Then create a circular mask to select only the desired well. Then, use that mask to apply the colour histogram operation to that well.

Your masked image should look like this:

Masked well plate

And, the program should produce a colour histogram that looks like this:

Well plate histogram

Summary Quiz#