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Dentate gyrus dataset

This tutorial uses raw data from scvelo package. Special thanks go to the scvelo team!

[ ]:

# get the latest version from pypi
# for other installations approaches, see https://dynamo-release.readthedocs.io/en/latest/ten_minutes_to_dynamo.html#how-to-install
!pip install dynamo-release --upgrade --quiet

[1]:

# from IPython.core.display import display, HTML
# display(HTML("<style>.container { width:90% !important; }</style>"))
# %matplotlib inline

import warnings
warnings.filterwarnings('ignore')

import dynamo as dyn

dyn.get_all_dependencies_version()
package dynamo-release umap-learn anndata cvxopt hdbscan loompy matplotlib numba numpy pandas pynndescent python-igraph scikit-learn scipy seaborn setuptools statsmodels tqdm trimap numdifftools colorcet
version 0.95.2 0.4.6 0.7.4 1.2.3 0.8.26 3.0.6 3.3.0 0.51.0 1.19.1 1.1.1 0.4.8 0.8.2 0.23.2 1.5.2 0.9.0 49.6.0 0.11.1 4.48.2 1.0.12 0.9.39 2.0.2 
[2]:

# emulate ggplot2 plotting styple with white background
dyn.configuration.set_figure_params('dynamo', background='white')


[3]:

adata = dyn.sample_data.DentateGyrus_scvelo()

adata.obsm['X_umap_ori'] = adata.obsm['X_umap'].copy()

[4]:

dyn.pl.show_fraction(adata)
dyn.pl.basic_stats(adata)
_images/dentategyrus_subset_scvelo_5_0.png
_images/dentategyrus_subset_scvelo_5_1.png 
[5]:

dyn.pp.recipe_monocle(adata, n_top_genes=2000, fg_kwargs={'shared_count': 30})


[5]:

AnnData object with n_obs × n_vars = 2930 × 13913
    obs: 'clusters', 'age(days)', 'clusters_enlarged', 'nGenes', 'nCounts', 'pMito', 'use_for_pca', 'unspliced_Size_Factor', 'initial_unspliced_cell_size', 'Size_Factor', 'initial_cell_size', 'spliced_Size_Factor', 'initial_spliced_cell_size', 'ntr', 'cell_cycle_phase'
    var: 'pass_basic_filter', 'log_m', 'log_cv', 'score', 'use_for_pca', 'ntr'
    uns: 'clusters_colors', 'velocyto_SVR', 'pp_norm_method', 'PCs', 'explained_variance_ratio_', 'pca_fit', 'feature_selection'
    obsm: 'X_umap', 'X_umap_ori', 'X_pca', 'X', 'cell_cycle_scores'
    layers: 'ambiguous', 'spliced', 'unspliced', 'X_unspliced', 'X_spliced'

[6]:

dyn.pl.variance_explained(adata)
dyn.pl.feature_genes(adata)
_images/dentategyrus_subset_scvelo_7_0.png
_images/dentategyrus_subset_scvelo_7_1.png 
[7]:

dyn.tl.dynamics(adata, model='stochastic', cores=3)
# or dyn.tl.dynamics(adata, model='deterministic')
# or dyn.tl.dynamics(adata, model='stochastic', est_method='negbin')

[7]:

AnnData object with n_obs × n_vars = 2930 × 13913
    obs: 'clusters', 'age(days)', 'clusters_enlarged', 'nGenes', 'nCounts', 'pMito', 'use_for_pca', 'unspliced_Size_Factor', 'initial_unspliced_cell_size', 'Size_Factor', 'initial_cell_size', 'spliced_Size_Factor', 'initial_spliced_cell_size', 'ntr', 'cell_cycle_phase'
    var: 'pass_basic_filter', 'log_m', 'log_cv', 'score', 'use_for_pca', 'ntr', 'beta', 'gamma', 'half_life', 'alpha_b', 'alpha_r2', 'gamma_b', 'gamma_r2', 'gamma_logLL', 'delta_b', 'delta_r2', 'uu0', 'ul0', 'su0', 'sl0', 'U0', 'S0', 'total0', 'use_for_dynamics'
    uns: 'clusters_colors', 'velocyto_SVR', 'pp_norm_method', 'PCs', 'explained_variance_ratio_', 'pca_fit', 'feature_selection', 'dynamics'
    obsm: 'X_umap', 'X_umap_ori', 'X_pca', 'X', 'cell_cycle_scores'
    layers: 'ambiguous', 'spliced', 'unspliced', 'X_unspliced', 'X_spliced', 'M_u', 'M_uu', 'M_s', 'M_us', 'M_ss', 'velocity_S'
    obsp: 'moments_con'

[8]:

# enforce recalculating the umap embedding. By default dynamo will avoid recalculation if a reduced dimension space exists.
dyn.tl.reduceDimension(adata, enforce=True)


[8]:

AnnData object with n_obs × n_vars = 2930 × 13913
    obs: 'clusters', 'age(days)', 'clusters_enlarged', 'nGenes', 'nCounts', 'pMito', 'use_for_pca', 'unspliced_Size_Factor', 'initial_unspliced_cell_size', 'Size_Factor', 'initial_cell_size', 'spliced_Size_Factor', 'initial_spliced_cell_size', 'ntr', 'cell_cycle_phase'
    var: 'pass_basic_filter', 'log_m', 'log_cv', 'score', 'use_for_pca', 'ntr', 'beta', 'gamma', 'half_life', 'alpha_b', 'alpha_r2', 'gamma_b', 'gamma_r2', 'gamma_logLL', 'delta_b', 'delta_r2', 'uu0', 'ul0', 'su0', 'sl0', 'U0', 'S0', 'total0', 'use_for_dynamics'
    uns: 'clusters_colors', 'velocyto_SVR', 'pp_norm_method', 'PCs', 'explained_variance_ratio_', 'pca_fit', 'feature_selection', 'dynamics', 'neighbors', 'umap_fit'
    obsm: 'X_umap', 'X_umap_ori', 'X_pca', 'X', 'cell_cycle_scores'
    layers: 'ambiguous', 'spliced', 'unspliced', 'X_unspliced', 'X_spliced', 'M_u', 'M_uu', 'M_s', 'M_us', 'M_ss', 'velocity_S'
    obsp: 'moments_con', 'connectivities', 'distances'

[9]:

dyn.tl.cell_velocities(adata)


calculating transition matrix via pearson kernel with sqrt transform.: 100%|██████████| 2930/2930 [00:05<00:00, 498.05it/s]
projecting velocity vector to low dimensional embedding...: 100%|██████████| 2930/2930 [00:00<00:00, 3557.37it/s]

[9]:

AnnData object with n_obs × n_vars = 2930 × 13913
    obs: 'clusters', 'age(days)', 'clusters_enlarged', 'nGenes', 'nCounts', 'pMito', 'use_for_pca', 'unspliced_Size_Factor', 'initial_unspliced_cell_size', 'Size_Factor', 'initial_cell_size', 'spliced_Size_Factor', 'initial_spliced_cell_size', 'ntr', 'cell_cycle_phase'
    var: 'pass_basic_filter', 'log_m', 'log_cv', 'score', 'use_for_pca', 'ntr', 'beta', 'gamma', 'half_life', 'alpha_b', 'alpha_r2', 'gamma_b', 'gamma_r2', 'gamma_logLL', 'delta_b', 'delta_r2', 'uu0', 'ul0', 'su0', 'sl0', 'U0', 'S0', 'total0', 'use_for_dynamics', 'use_for_transition'
    uns: 'clusters_colors', 'velocyto_SVR', 'pp_norm_method', 'PCs', 'explained_variance_ratio_', 'pca_fit', 'feature_selection', 'dynamics', 'neighbors', 'umap_fit', 'grid_velocity_umap'
    obsm: 'X_umap', 'X_umap_ori', 'X_pca', 'X', 'cell_cycle_scores', 'velocity_umap'
    layers: 'ambiguous', 'spliced', 'unspliced', 'X_unspliced', 'X_spliced', 'M_u', 'M_uu', 'M_s', 'M_us', 'M_ss', 'velocity_S'
    obsp: 'moments_con', 'connectivities', 'distances', 'pearson_transition_matrix'

[10]:

DentateGyrus_genes = ["Tnc", "Gfap", "Tac2", "Pdgfra", "Igfbpl1", 'Ptprn', "Sema3c", "Neurod6", "Stmn2", "Sema5a", "C1ql3", "Cpne4", "Cck"]

dyn.pl.phase_portraits(adata, genes=DentateGyrus_genes, ncols=3, figsize=(3, 3), basis='pca', show_quiver=False)
_images/dentategyrus_subset_scvelo_11_0.png 
[11]:

dyn.pl.cell_wise_vectors(adata, color=['clusters'], quiver_size=6, quiver_length=6, figsize=(6, 5), pointsize=0.1) # ['GRIA3', 'LINC00982', 'AFF2']



<Figure size 600x500 with 0 Axes>
_images/dentategyrus_subset_scvelo_12_1.png 
[12]:

dyn.pl.grid_vectors(adata, color=['clusters'], method='gaussian')

<Figure size 600x400 with 0 Axes>
_images/dentategyrus_subset_scvelo_13_1.png 
[13]:

dyn.pl.streamline_plot(adata, color=['clusters'], basis='umap', density=1, background='white',s_kwargs_dict={"alpha": 0.05})

<Figure size 600x400 with 0 Axes>
_images/dentategyrus_subset_scvelo_14_1.png 
[14]:

dyn.pl.streamline_plot(adata, color=['clusters'], basis='umap_ori', density=1, background='white',s_kwargs_dict={"alpha": 0.05})

projecting velocity vector to low dimensional embedding...:  27%|██▋       | 784/2930 [00:00<00:00, 3801.14it/s]

Using existing pearson_transition_matrix found in .obsp.

projecting velocity vector to low dimensional embedding...: 100%|██████████| 2930/2930 [00:00<00:00, 4048.10it/s]

<Figure size 600x400 with 0 Axes>
_images/dentategyrus_subset_scvelo_15_4.png 
[15]:

# you can set `verbose = 1/2/3` to obtain different levels of running information of vector field reconstruction
dyn.vf.VectorField(adata, basis='umap_ori', dims=[0, 1])


[16]:

dyn.pl.topography(adata, color=['clusters', 'Tnc'], basis='umap_ori', ncols=2)
_images/dentategyrus_subset_scvelo_17_0.png 
[17]:

dyn.vf.VectorField(adata, basis='umap_ori', dims=[0, 1], pot_curl_div=True)


[18]:

dyn.ext.ddhodge(adata, basis='umap_ori')

Constructing diffusion graph from reconstructed vector field: 2930it [01:57, 24.83it/s]

[19]:

adata

[19]:

AnnData object with n_obs × n_vars = 2930 × 13913
    obs: 'clusters', 'age(days)', 'clusters_enlarged', 'nGenes', 'nCounts', 'pMito', 'use_for_pca', 'unspliced_Size_Factor', 'initial_unspliced_cell_size', 'Size_Factor', 'initial_cell_size', 'spliced_Size_Factor', 'initial_spliced_cell_size', 'ntr', 'cell_cycle_phase', 'umap_ori_ddhodge_div', 'umap_ori_ddhodge_potential'
    var: 'pass_basic_filter', 'log_m', 'log_cv', 'score', 'use_for_pca', 'ntr', 'beta', 'gamma', 'half_life', 'alpha_b', 'alpha_r2', 'gamma_b', 'gamma_r2', 'gamma_logLL', 'delta_b', 'delta_r2', 'uu0', 'ul0', 'su0', 'sl0', 'U0', 'S0', 'total0', 'use_for_dynamics', 'use_for_transition'
    uns: 'clusters_colors', 'velocyto_SVR', 'pp_norm_method', 'PCs', 'explained_variance_ratio_', 'pca_fit', 'feature_selection', 'dynamics', 'neighbors', 'umap_fit', 'grid_velocity_umap', 'grid_velocity_umap_ori', 'VecFld_umap_ori', 'VecFld'
    obsm: 'X_umap', 'X_umap_ori', 'X_pca', 'X', 'cell_cycle_scores', 'velocity_umap', 'velocity_umap_ori', 'velocity_umap_ori_SparseVFC', 'X_umap_ori_SparseVFC'
    layers: 'ambiguous', 'spliced', 'unspliced', 'X_unspliced', 'X_spliced', 'M_u', 'M_uu', 'M_s', 'M_us', 'M_ss', 'velocity_S'
    obsp: 'moments_con', 'connectivities', 'distances', 'pearson_transition_matrix', 'umap_ori_ddhodge'

[20]:

dyn.pl.topography(adata, color=['clusters', 'umap_ori_ddhodge_div', 'umap_ori_ddhodge_potential'],
                  basis='umap_ori', ncols=3, frontier=True)
_images/dentategyrus_subset_scvelo_21_0.png 
[21]:

# emulate ggplot2 plotting styple with black background
dyn.configuration.set_figure_params('dynamo', background='black')

[22]:


dyn.pl.phase_portraits(adata, genes=DentateGyrus_genes, ncols=3, figsize=(6, 4), basis='umap_ori', show_quiver=False)

dyn.pl.cell_wise_vectors(adata, color=['clusters'], basis='umap_ori', pointsize=0.1, quiver_size=4, quiver_length=4, background='black') # ['GRIA3', 'LINC00982', 'AFF2']

dyn.pl.grid_vectors(adata, color=['clusters'], basis='umap_ori', method='gaussian', background='black')

dyn.pl.streamline_plot(adata, color=['clusters'], basis='umap_ori', density=2, background='black')

dyn.pl.topography(adata, color=['clusters', 'Tnc'], basis='umap_ori', ncols=2, background='black')
_images/dentategyrus_subset_scvelo_23_0.png 
<Figure size 600x400 with 0 Axes>
_images/dentategyrus_subset_scvelo_23_2.png 
<Figure size 600x400 with 0 Axes>
_images/dentategyrus_subset_scvelo_23_4.png 
<Figure size 600x400 with 0 Axes>
_images/dentategyrus_subset_scvelo_23_6.png
_images/dentategyrus_subset_scvelo_23_7.png 
[23]:

progenitor = adata.obs_names[adata.obs.clusters.isin(['nIPC', 'Neuroblast'])]
len(progenitor)

[23]:

436

[24]:

dyn.pd.fate(adata, basis='umap_ori', init_cells=progenitor, interpolation_num=100,  direction='forward',
   inverse_transform=False, average=False, cores=3)

[24]:

AnnData object with n_obs × n_vars = 2930 × 13913
    obs: 'clusters', 'age(days)', 'clusters_enlarged', 'nGenes', 'nCounts', 'pMito', 'use_for_pca', 'unspliced_Size_Factor', 'initial_unspliced_cell_size', 'Size_Factor', 'initial_cell_size', 'spliced_Size_Factor', 'initial_spliced_cell_size', 'ntr', 'cell_cycle_phase', 'umap_ori_ddhodge_div', 'umap_ori_ddhodge_potential'
    var: 'pass_basic_filter', 'log_m', 'log_cv', 'score', 'use_for_pca', 'ntr', 'beta', 'gamma', 'half_life', 'alpha_b', 'alpha_r2', 'gamma_b', 'gamma_r2', 'gamma_logLL', 'delta_b', 'delta_r2', 'uu0', 'ul0', 'su0', 'sl0', 'U0', 'S0', 'total0', 'use_for_dynamics', 'use_for_transition'
    uns: 'clusters_colors', 'velocyto_SVR', 'pp_norm_method', 'PCs', 'explained_variance_ratio_', 'pca_fit', 'feature_selection', 'dynamics', 'neighbors', 'umap_fit', 'grid_velocity_umap', 'grid_velocity_umap_ori', 'VecFld_umap_ori', 'VecFld', 'fate_umap_ori'
    obsm: 'X_umap', 'X_umap_ori', 'X_pca', 'X', 'cell_cycle_scores', 'velocity_umap', 'velocity_umap_ori', 'velocity_umap_ori_SparseVFC', 'X_umap_ori_SparseVFC'
    layers: 'ambiguous', 'spliced', 'unspliced', 'X_unspliced', 'X_spliced', 'M_u', 'M_uu', 'M_s', 'M_us', 'M_ss', 'velocity_S'
    obsp: 'moments_con', 'connectivities', 'distances', 'pearson_transition_matrix', 'umap_ori_ddhodge'

[25]:

%%capture
import matplotlib.pyplot as plt

fig, ax = plt.subplots()
ax = dyn.pl.topography(adata, color='clusters', ax=ax, save_show_or_return='return', basis='umap_ori')

[26]:

%%capture
instance = dyn.mv.StreamFuncAnim(adata=adata, ax=ax, basis='umap_ori', color='clusters')

[27]:

from matplotlib import animation
import numpy as np

anim = animation.FuncAnimation(instance.fig, instance.update, init_func=instance.init_background,
                                frames=np.arange(100), interval=100, blit=True)
from IPython.core.display import display, HTML
HTML(anim.to_jshtml()) # embedding to jupyter notebook.

[27]:

[28]:

%%capture
fig, ax = plt.subplots()
ax = dyn.pl.topography(adata, color='clusters', basis='umap_ori', ax=ax, save_show_or_return='return')
dyn.mv.animate_fates(adata, color='clusters', basis='umap_ori', n_steps=100, fig=fig, ax=ax,
                     save_show_or_return='save', logspace=True, max_time=None, save_kwargs={"filename": 'dentategyrus.gif'})