Please send feedback and report any problems to the [Photon-HDF5 google group](https://groups.google.com/forum/#!forum/photon-hdf5).
The notebook is composed by "text cells", such as this paragraph, and "code cells"
containing the code to be executed (and identified by an
In [ ] prompt).
To execute a code cell, select it and press SHIFT+ENTER.
To modify an cell, click on it to enter "edit mode" (indicated by a green frame),
You can run this notebook directly online (for demo purposes), or you can run it on your on desktop. For a local installation please refer to:
Before starting, you have to upload a data file to be converted to Photon-HDF5. You can use one of our example data files available on figshare.
To upload a file (up to 35 MB) switch to the "Home" tab in your browser, click the Upload button and select the data file and wait until the upload completes. To upload files larger than 35 MB (like some of our example files) please use the Upload notebook instead.
Once the file is uploaded, come back here and follow the instructions below.
Specify the input data file in the following cell:
filename = 'Pre.ht3'
The next cell will check if the
filename location is correct:
import os try: with open(filename): pass print('Data file found, you can proceed.') except IOError: print('ATTENTION: Data file not found, please check the filename.\n' ' (current value "%s")' % filename)
In case of file not found, please double check the file name and that the file has been uploaded.
%matplotlib inline import numpy as np import phconvert as phc print('phconvert version: ' + phc.__version__)
Then we load the input file:
d, meta = phc.loader.nsalex_ht3(filename, donor = 1, acceptor = 0, alex_period_donor = (1540, 3050), alex_period_acceptor = (150, 1500), excitation_wavelengths = (470e-9, 635e-9), detection_wavelengths = (525e-9, 690e-9), )
And we plot the
The previous plot is the
nanotimes histogram for the donor and acceptor channel separately.
The shaded areas marks the donor (green) and acceptor (red) excitation periods.
If the histogram looks wrong in some aspects (no photons, wrong detectors assignment, wrong period selection) please go back to the previous cell which loads the file and change the parameters until the histogram looks correct.
You may also find useful to see how many different detectors are present and their number of photons. This information is shown in the next cell:
detectors = d['photon_data']['detectors'] print("Detector Counts") print("-------- --------") for det, count in zip(*np.unique(detectors, return_counts=True)): print("%8d %8d" % (det, count))
In the next few cells, we specify some metadata that will be stored in the Photon-HDF5 file. Please modify these fields to reflect the content of the data file:
author = 'John Doe' author_affiliation = 'Research Institution' description = 'A demonstrative smFRET-nsALEX measurement.' sample_name = 'Doubly-labeled ssDNA partially hybridized to a complementary strand.' dye_names = 'ATTO488, ATTO647N' buffer_name = 'Tris20 mM Ph 7.8'
Once you finished editing the the previous sections you can proceed with the actual conversion. To do that, click on the menu Cells -> Run All Below.
After the execution go to Section 6 to download the Photon-HDF5 file.
The cells below contain the code to convert the input file to Photon-HDF5.
d['description'] = description d['sample'] = dict( sample_name=sample_name, dye_names=dye_names, buffer_name=buffer_name, num_dyes = len(dye_names.split(','))) d['identity'] = dict( author=author, author_affiliation=author_affiliation)
This command saves the new file to disk. If the input data does not follows the Photon-HDF5 specification it returns an error (
Invalid_PhotonHDF5) printing what violates the specs.
from pprint import pprint
filename = d['_data_file'].filename
h5data = phc.hdf5.load_photon_hdf5(filename)