Storing Image Data in NWB

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:
Table of Contents
Create an NWB File OpticalSeries: Storing series of images as stimuli ImageSeries: Storing series of images as acquisition External Files Static Images RGBAImage: for color images with transparency RGBImage: for color images GrayscaleImage: for grayscale images Images: a container for images Index Series for Repeated Images Writing the images to an NWB File

Create an NWB File

nwb = NwbFile( ...
'session_description', 'mouse in open exploration',...
'identifier', 'Mouse5_Day3', ...
'session_start_time', datetime(2018, 4, 25, 2, 30, 3, 'TimeZone', 'local'), ...
'timestamps_reference_time', datetime(2018, 4, 25, 3, 0, 45, 'TimeZone', 'local'), ...
'general_experimenter', 'LastName, FirstName', ... % optional
'general_session_id', 'session_1234', ... % optional
'general_institution', 'University of My Institution', ... % optional
'general_related_publications', 'DOI:10.1016/j.neuron.2016.12.011' ... % optional
);
nwb
nwb =
NwbFile with properties: nwb_version: '2.6.0' file_create_date: [] identifier: 'Mouse5_Day3' session_description: 'mouse in open exploration' session_start_time: {[2018-04-25T02:30:03.000000+02:00]} timestamps_reference_time: {[2018-04-25T03:00:45.000000+02:00]} acquisition: [0×1 types.untyped.Set] analysis: [0×1 types.untyped.Set] general: [0×1 types.untyped.Set] general_data_collection: '' general_devices: [0×1 types.untyped.Set] general_experiment_description: '' general_experimenter: 'LastName, FirstName' general_extracellular_ephys: [0×1 types.untyped.Set] general_extracellular_ephys_electrodes: [] general_institution: 'University of My Institution' general_intracellular_ephys: [0×1 types.untyped.Set] general_intracellular_ephys_experimental_conditions: [] general_intracellular_ephys_filtering: '' general_intracellular_ephys_intracellular_recordings: [] general_intracellular_ephys_repetitions: [] general_intracellular_ephys_sequential_recordings: [] general_intracellular_ephys_simultaneous_recordings: [] general_intracellular_ephys_sweep_table: [] general_keywords: '' general_lab: '' general_notes: '' general_optogenetics: [0×1 types.untyped.Set] general_optophysiology: [0×1 types.untyped.Set] general_pharmacology: '' general_protocol: '' general_related_publications: 'DOI:10.1016/j.neuron.2016.12.011' general_session_id: 'session_1234' general_slices: '' general_source_script: '' general_source_script_file_name: '' general_stimulus: '' general_subject: [] general_surgery: '' general_virus: '' intervals: [0×1 types.untyped.Set] intervals_epochs: [] intervals_invalid_times: [] intervals_trials: [] processing: [0×1 types.untyped.Set] scratch: [0×1 types.untyped.Set] stimulus_presentation: [0×1 types.untyped.Set] stimulus_templates: [0×1 types.untyped.Set] units: []

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 ('RGB', 'y', 'x', 'time') = (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.
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.
image_data = randi(255, [3, 50, 50, 200]);
optical_series = types.core.OpticalSeries( ...
'distance', 0.7, ... % required
'field_of_view', [0.2, 0.3, 0.7], ... % required
'orientation', 'lower left', ... % required
'data', image_data, ...
'data_unit', 'n.a.', ...
'starting_time_rate', 1.0, ...
'starting_time', 0.0, ...
'description', 'The images presented to the subject as stimuli' ...
);
 
nwb.stimulus_presentation.set('StimulusPresentation', 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.
image_data = randi(255, [3, 50, 50, 200]);
behavior_images = types.core.ImageSeries( ...
'data', image_data, ...
'description', 'Image data of an animal in environment', ...
'data_unit', 'n.a.', ...
'starting_time_rate', 1.0, ...
'starting_time', 0.0 ...
);
 
nwb.acquisition.set('ImageSeries', 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. external_file can be a cell array of multiple video files.
The starting_frame attribute serves as an index to indicate the starting frame of each external file, allowing you to skip the beginning of videos.
external_files = {'video1.pmp4', 'video2.pmp4'};
 
timestamps = [0.0, 0.04, 0.07, 0.1, 0.14, 0.16, 0.21];
behavior_external_file = types.core.ImageSeries( ...
'description', 'Behavior video of animal moving in environment', ...
'data_unit', 'n.a.', ...
'external_file', external_files, ...
'format', 'external', ...
'external_file_starting_frame', [0, 2, 4], ...
'timestamps', timestamps ...
);
 
nwb.acquisition.set('ExternalVideos', behavior_external_file);

Static Images

Static images can be stored in an NWBFile object by creating an RGBAImage, RGBImage or GrayscaleImage object with the image data. All of these image types provide an optional description parameter to include text description about the image and the 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.
image_data = randi(255, [4, 200, 200]);
 
rgba_image = types.core.RGBAImage( ...
'data', image_data, ... % required
'resolution', 70.0, ...
'description', 'RGBA image' ...
);

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.
image_data = randi(255, [3, 200, 200]);
 
rgb_image = types.core.RGBImage( ...
'data', image_data, ... % required
'resolution', 70.0, ...
'description', 'RGB image' ...
);

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.
image_data = randi(255, [200, 200]);
 
grayscale_image = types.core.GrayscaleImage( ...
'data', image_data, ... % required
'resolution', 70.0, ...
'description', 'Grayscale image' ...
);

Images: a container for images

Add the images to an Images container that accepts any of these image types.
image_collection = types.core.Images( ...
'description', 'A collection of logo images presented to the subject.'...
);
 
image_collection.image.set('rgba_image', rgba_image);
image_collection.image.set('rgb_image', rgb_image);
image_collection.image.set('grayscale_image', grayscale_image);
 
nwb.acquisition.set('image_collection', image_collection);

Index Series for Repeated Images

You may want to set up a time series of images where some images are repeated many times. You could create an ImageSeries that repeats the data each time the image is shown, but that would be inefficient, because it would store the same data multiple times. A better solution would be to store the unique images once and reference those images. This is how IndexSeries works. First, create an Images container with the order of images defined using an ImageReferences. Then create an IndexSeries that indexes into the Images.
rgbImage = imread('street2.jpg');
grayImage = uint8(sum(double(rgbImage), 3) ./ double(max(max(max(rgbImage)))));
GsStreet = types.core.GrayscaleImage(...
'data', grayImage, ...
'description', 'grayscale image of a street.', ...
'resolution', 28 ...
);
 
RgbStreet = types.core.RGBImage( ...
'data', rgbImage, ...
'resolution', 28, ...
'description', 'RGB Street' ...
);
 
ImageOrder = types.core.ImageReferences(...
'data', [types.untyped.ObjectView(RgbStreet), types.untyped.ObjectView(GsStreet)] ...
);
Images = types.core.Images( ...
'gs_face', GsStreet, ...
'rgb_face', RgbStreet, ...
'description', 'A collection of streets.', ...
'order_of_images', ImageOrder ...
);
 
types.core.IndexSeries(...
'data', [0, 1, 0, 1], ... % NOTE: 0-indexed
'indexed_images', Images, ...
'timestamps', [0.1, 0.2, 0.3, 0.4] ...
)
ans =
IndexSeries with properties: indexed_images: [1×1 types.core.Images] indexed_timeseries: [] starting_time_unit: 'seconds' timestamps_interval: 1 timestamps_unit: 'seconds' data: [0 1 0 1] comments: 'no comments' control: [] control_description: '' data_continuity: '' data_conversion: [] data_offset: [] data_resolution: [] data_unit: 'N/A' description: 'no description' starting_time: [] starting_time_rate: [] timestamps: [0.1000 0.2000 0.3000 0.4000]
Here data contains the (0-indexed) index of the displayed image as they are ordered in the ImageReference.

Writing the images to an NWB File

Now use nwbExport to write the file.
nwbExport(nwb, "images_test.nwb");