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d3eee8e
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Parent(s):
0dab632
Add scipy requirement
Browse files- analyse.py +0 -157
- app.py +6 -3
- requirements.txt +1 -0
analyse.py
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import lxml.etree as ET
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import gzip
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import tifffile
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import matplotlib.pyplot as plt
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import numpy as np
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from PIL import Image, ImageDraw
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import pandas as pd
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def get_paths_from_traces_file(traces_file):
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tree = ET.parse(traces_file)
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root = tree.getroot()
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all_paths = []
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path_lengths = []
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for path in root.findall('path'):
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length=path.get('reallength')
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path_points = []
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for point in path:
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path_points.append((int(point.get('x')), int(point.get('y')), int(point.get('z'))))
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all_paths.append(path_points)
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path_lengths.append(length)
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return all_paths, path_lengths
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def visualise_ordering(points_list, dim):
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rdim, cdim, _ = dim
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vis = np.zeros((rdim, cdim, 3), dtype=np.uint8)
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def get_col(i):
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r = int(255 * i/len(points_list))
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g = 255 - r
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return r, g, 0
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for n, p in enumerate(points_list):
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c, r, _ = p
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wr, wc = 5, 5
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vis[max(0,r-wr):min(rdim,r+wr),max(0,c-wc):min(cdim,c+wc)] = get_col(n)
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return vis
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col_map = [(255,0,0), (0,255,0), (0,0,255), (255,255,0), (255,0,255), (0,255,255)]
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def draw_paths(all_paths, foci_stack):
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im = np.max(foci_stack, axis=0)
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im = (im/np.max(im)*255).astype(np.uint8)
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im = np.dstack((im,)*3)
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im = Image.fromarray(im) #.convert('RGB')
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draw = ImageDraw.Draw(im)
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for i, (p, col) in enumerate(zip(all_paths, col_map)):
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draw.line([(u[0], u[1]) for u in p], fill=col)
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draw.text((p[0][0], p[0][1]), str(i+1), fill=col)
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return im
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# Sum of measure_stack over regin where mask==1
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def measure_from_mask(mask, measure_stack):
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return np.sum(mask * measure_stack)
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# Max of measure_stack over region where mask==1
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def max_from_mask(mask, measure_stack):
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return np.max(mask * measure_stack)
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# Translate mask to point p, treating makss near stack edges correctly
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def make_mask_s(p, melem, measure_stack):
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mask = melem
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R = melem.shape[0] // 2
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r, c, z = p
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m_data = np.zeros(melem.shape)
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s = measure_stack.shape
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o_1, o_2, o_3 = max(R-r, 0), max(R-c, 0), max(R-z,0)
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e_1, e_2, e_3 = min(R-r+s[0], 2*R), min(R-c+s[1], 2*R), min(R-z+s[2], 2*R)
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m_data[o_1:e_1,o_2:e_2,o_3:e_3] = measure_stack[max(r-R,0):min(r+R,s[0]),max(c-R,0):min(c+R,s[1]),max(z-R,0):min(z+R, s[2])]
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return mask, m_data
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# Measure the (mean/max) value of measure_stack about the point p, using
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# the structuring element melem. op indicates the appropriate measurement (mean/max)
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def measure_at_point(p, melem, measure_stack, op='mean'):
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if op=='mean':
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mask, m_data = make_mask_s(p, melem, measure_stack)
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melem_size = np.sum(melem)
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return float(measure_from_mask(mask, m_data) / melem_size)
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else:
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mask, m_data = make_mask_s(p, melem, measure_stack)
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return float(max_from_mask(mask, m_data))
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# Generate spherical region
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def make_sphere(R=5, z_scale_ratio=2.3):
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x, y, z = np.ogrid[-R:R, -R:R, -R:R]
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sphere = x**2 + y**2 + (z_scale_ratio * z)**2 < R**2
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return sphere
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# Measure the values of measure_stack at each of the points of points_list in turn.
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# Measurement is the mean / max (specified by op) on the spherical region about each point
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def measure_all_with_sphere(points_list, measure_stack, op='mean'):
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melem = make_sphere()
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measure_func = lambda p: measure_at_point(p, melem, measure_stack, op)
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return list(map(measure_func, points_list))
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# Measure fluorescence levels along ordered skeleton
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def measure_chrom2(path, hei10):
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# single chrom - structure containing skeleton (single_chrom.skel) and
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# fluorecence levels (single_chrom.hei10) as Image3D objects (equivalent to ndarray)
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# Returns list of coordinates in skeleton, the ordered path
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vis = visualise_ordering(path, dim=hei10.shape)
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measurements = measure_all_with_sphere(path, hei10, op='mean')
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measurements_max = measure_all_with_sphere(path, hei10, op='max')
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return vis, measurements, measurements_max
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def extract_peaks(cell_id, all_paths, path_lengths, measured_traces):
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n = len(all_paths)
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#headers = ['Cell_ID', 'Trace', 'Trace_length(um)', 'detection_sphere_radius(um)', 'Foci_ID_threshold', 'Foci_per_trace']
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#for i in range(max_n):
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# headers += [f'Foci{i}_relative_intensity', f'Foci_{i}_position(um)']
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data_dict = {}
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data_dict['Cell_ID'] = [cell_id]*n
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data_dict['Trace'] = range(1, n+1)
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data_dict['Trace_length(um)'] = path_lengths
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data_dict['Detection_sphere_radius(um)'] = [0.2]*n
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data_dict['Foci_ID_threshold'] = [0.4]*n
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return pd.DataFrame(data_dict)
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def analyse_paths(cell_id, foci_file, traces_file):
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foci_stack = tifffile.imread(foci_file)
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all_paths, path_lengths = get_paths_from_traces_file(traces_file)
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all_trace_vis = []
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all_m = []
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for p in all_paths:
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vis, m, _ = measure_chrom2(p,foci_stack.transpose(2,1,0))
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all_trace_vis.append(vis)
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all_m.append(m)
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trace_overlay = draw_paths(all_paths, foci_stack)
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fig, ax = plt.subplots(len(all_paths),1)
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for i, m in enumerate(all_m):
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ax[i].plot(m)
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extracted_peaks = extract_peaks(cell_id, all_paths, path_lengths, all_m)
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return trace_overlay, all_trace_vis, fig, extracted_peaks
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app.py
CHANGED
@@ -40,6 +40,7 @@ with gr.Blocks() as demo:
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# Resolutions for xy and z axis
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threshold_type = gr.Radio(["per-trace", "per-cell"], label="Threshold-type", value="per-trace", interactive=True)
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# The output column showing the result of processing
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data_file_output=gr.File(label="Output data file (.csv)")
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def process(cellid_input, image_input, path_input, sphere_radius, peak_threshold, xy_res, z_res, threshold_type):
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config = { 'sphere_radius': sphere_radius,
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'peak_threshold': peak_threshold,
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'xy_res': xy_res,
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'z_res': z_res,
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'threshold_type': threshold_type
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paths, traces, fig, extracted_peaks = analyse_paths(cellid_input, image_input.name, path_input.name, config)
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with gr.Row():
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greet_btn = gr.Button("Process")
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greet_btn.click(fn=process, inputs=[cellid_input, image_input, path_input, sphere_radius, peak_threshold, xy_res, z_res, threshold_type], outputs=[trace_output, image_output, plot_output, data_output, data_file_output], api_name="process")
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if __name__ == "__main__":
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# Resolutions for xy and z axis
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threshold_type = gr.Radio(["per-trace", "per-cell"], label="Threshold-type", value="per-trace", interactive=True)
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use_corrected_positions = gr.Checkbox(label="Correct foci position measurements", value=True, interactive=True)
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# The output column showing the result of processing
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data_file_output=gr.File(label="Output data file (.csv)")
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def process(cellid_input, image_input, path_input, sphere_radius, peak_threshold, xy_res, z_res, threshold_type, use_corrected_positions):
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config = { 'sphere_radius': sphere_radius,
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'peak_threshold': peak_threshold,
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'xy_res': xy_res,
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'z_res': z_res,
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'threshold_type': threshold_type,
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'use_corrected_positions': use_corrected_positions
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}
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paths, traces, fig, extracted_peaks = analyse_paths(cellid_input, image_input.name, path_input.name, config)
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with gr.Row():
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greet_btn = gr.Button("Process")
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greet_btn.click(fn=process, inputs=[cellid_input, image_input, path_input, sphere_radius, peak_threshold, xy_res, z_res, threshold_type, use_corrected_positions], outputs=[trace_output, image_output, plot_output, data_output, data_file_output], api_name="process")
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if __name__ == "__main__":
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requirements.txt
CHANGED
@@ -4,3 +4,4 @@ matplotlib
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numpy
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lxml
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pandas
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numpy
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lxml
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pandas
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scipy
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