Storing Image Data in NWB

This tutorial will demonstrate the usage of the pynwb.image module for adding images to an NWBFile.

Image data can be a collection of individual images or movie segments (as a movie is simply a series of images), about the subject, the environment, the presented stimuli, or other parts related to the experiment. This tutorial focuses in particular on the usage of:

• OpticalSeries for series of images that were presented as stimulus

• ImageSeries, for series of images (movie segments);

• GrayscaleImage, RGBImage, RGBAImage, for static images;

The following examples will reference variables that may not be defined within the block they are used in. For clarity, we define them here:

from datetime import datetime
from dateutil import tz

import numpy as np
from PIL import Image

from pynwb import NWBFile, NWBHDF5IO
from pynwb.base import Images
from pynwb.image import RGBAImage, RGBImage, GrayscaleImage, OpticalSeries, ImageSeries

# Define file paths used in the tutorial
import os
nwbfile_path = os.path.abspath("images_tutorial.nwb")
moviefiles_path = [
    os.path.abspath("image/file_1.tiff"),
    os.path.abspath("image/file_2.tiff"),
    os.path.abspath("image/file_3.tiff"),
]

Create an NWB File

Create an NWBFile object with the required fields (session_description, identifier, session_start_time) and additional metadata.

session_start_time = datetime(2018, 4, 25, 2, 30, 3, tzinfo=tz.gettz("US/Pacific"))

nwbfile = NWBFile(
    session_description="Mouse exploring an open field",  # required
    identifier="Mouse5_Day3",  # required
    session_start_time=session_start_time,  # required
    session_id="session_1234",  # optional
    experimenter="My Name",  # optional
    lab="My Lab Name",  # optional
    institution="University of My Institution",  # optional
    related_publications="DOI:10.1016/j.neuron.2016.12.011",  # optional
)

nwbfile

See also

You can learn more about the NWBFile format in the NWB File Basics tutorial.

OpticalSeries: Storing series of images as stimuli

OpticalSeries is for time series of images that were presented to the subject as stimuli. We will create an OpticalSeries object with the name "StimulusPresentation" representing what images were shown to the subject and at what times.

Image data can be stored either in the HDF5 file or as an external image file. For this tutorial, we will use fake image data with shape of ('time', 'x', 'y', 'RGB') = (200, 50, 50, 3). As in all TimeSeries, the first dimension is time. The second and third dimensions represent x and y. The fourth dimension represents the RGB value (length of 3) for color images.

NWB differentiates between acquired data and data that was presented as stimulus. We can add it to the NWBFile object as stimulus data using the add_stimulus method.

image_data = np.random.randint(low=0, high=255, size=(200, 50, 50, 3), dtype=np.uint8)
optical_series = OpticalSeries(
    name="StimulusPresentation",  # required
    distance=0.7,  # required
    field_of_view=[0.2, 0.3, 0.7],  # required
    orientation="lower left",  # required
    data=image_data,
    unit="n.a.",
    format="raw",
    starting_frame=[0.0],
    rate=1.0,
    comments="no comments",
    description="The images presented to the subject as stimuli",
)

nwbfile.add_stimulus(timeseries=optical_series)

ImageSeries: Storing series of images as acquisition

ImageSeries is a general container for time series of images acquired during the experiment. Image data can be stored either in the HDF5 file or as an external image file. When color images are stored in the HDF5 file the color channel order is expected to be RGB.

We can add raw data to the NWBFile object as acquisition using the add_acquisition method.

image_data = np.random.randint(low=0, high=255, size=(200, 50, 50, 3), dtype=np.uint8)
behavior_images = ImageSeries(
    name="ImageSeries",
    data=image_data,
    description="Image data of an animal moving in environment.",
    unit="n.a.",
    format="raw",
    rate=1.0,
    starting_time=0.0,
)

nwbfile.add_acquisition(behavior_images)

External Files

External files (e.g. video files of the behaving animal) can be added to the NWBFile by creating an ImageSeries object using the external_file attribute that specifies the path to the external file(s) on disk. The file(s) path must be relative to the path of the NWB file. Either external_file or data must be specified, but not both.

If the sampling rate is constant, use rate and starting_time to specify time. For irregularly sampled recordings, use timestamps to specify time for each sample image.

Each external image may contain one or more consecutive frames of the full ImageSeries. The starting_frame attribute serves as an index to indicate which frame each file contains. For example, if the external_file dataset has three paths to files and the first and the second file have 2 frames, and the third file has 3 frames, then this attribute will have values [0, 2, 4].

external_file = [
    os.path.relpath(movie_path, nwbfile_path) for movie_path in moviefiles_path
]
# We have 3 movie files each containing multiple frames. We here need to specify the timestamp for each frame.
timestamps = [0.0, 0.04, 0.07, 0.1, 0.14, 0.16, 0.21]
behavior_external_file = ImageSeries(
    name="ExternalFiles",
    description="Behavior video of animal moving in environment.",
    unit="n.a.",
    external_file=external_file,
    format="external",
    starting_frame=[0, 2, 4],
    timestamps=timestamps,
)

nwbfile.add_acquisition(behavior_external_file)

Note

See the External Links in NWB and DANDI guidelines of the DANDI data archive for best practices on how to organize external files, e.g., movies and images.

Static images

Static images can be stored in an NWBFile object by creating an RGBAImage, RGBImage or GrayscaleImage object with the image data.

Note

All basic image types RGBAImage, RGBImage, and GrayscaleImage provide the optional: 1) description parameter to include a text description about the image and 2) resolution parameter to specify the pixels / cm resolution of the image.

RGBAImage: for color images with transparency

RGBAImage is for storing data of color image with transparency. data must be 3D where the first and second dimensions represent x and y. The third dimension has length 4 and represents the RGBA value.

img = Image.open("docs/source/figures/logo_pynwb.png")  # an example image

rgba_logo = RGBAImage(
    name="pynwb_RGBA_logo",
    data=np.array(img),
    resolution=70.0,  # in pixels / cm
    description="RGBA version of the PyNWB logo.",
)

RGBImage: for color images

RGBImage is for storing data of RGB color image. data must be 3D where the first and second dimensions represent x and y. The third dimension has length 3 and represents the RGB value.

rgb_logo = RGBImage(
    name="pynwb_RGB_logo",
    data=np.array(img.convert("RGB")),
    resolution=70.0,
    description="RGB version of the PyNWB logo.",
)

GrayscaleImage: for grayscale images

GrayscaleImage is for storing grayscale image data. data must be 2D where the first and second dimensions represent x and y.

gs_logo = GrayscaleImage(
    name="pynwb_Grayscale_logo",
    data=np.array(img.convert("L")),
    description="Grayscale version of the PyNWB logo.",
    resolution=35.433071,
)

Images: a container for images

Add the images to an Images container that accepts any of these image types.

images = Images(
    name="logo_images",
    images=[rgb_logo, rgba_logo, gs_logo],
    description="A collection of logo images presented to the subject.",
)

nwbfile.add_acquisition(images)

Writing the images to an NWB File

As demonstrated in the Writing an NWB file tutorial, we will use NWBHDF5IO to write the file.

with NWBHDF5IO(nwbfile_path, "w") as io:
    io.write(nwbfile)

Reading and accessing data

To read the NWB file, use another NWBHDF5IO object, and use the read method to retrieve an NWBFile object.

We can access the data added as acquisition to the NWB File by indexing nwbfile.acquisition with the name of the ImageSeries object “ImageSeries”.

We can also access OpticalSeries data that was added to the NWB File as stimuli by indexing nwbfile.stimulus with the name of the OpticalSeries object “StimulusPresentation”. Data arrays are read passively from the file. Accessing the data attribute of the OpticalSeries object does not read the data values into memory, but returns an HDF5 object that can be indexed to read data. Use the [:] operator to read the entire data array into memory.

with NWBHDF5IO(nwbfile_path, "r") as io:
    read_nwbfile = io.read()
    print(read_nwbfile.acquisition["ImageSeries"])
    print(read_nwbfile.stimulus["StimulusPresentation"].data[:])