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from __future__ import annotations |
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import numpy as np |
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from monai.transforms.transform import Transform |
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class ExtractHEStains(Transform): |
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"""Class to extract a target stain from an image, using stain deconvolution (see Note). |
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Args: |
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tli: transmitted light intensity. Defaults to 240. |
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alpha: tolerance in percentile for the pseudo-min (alpha percentile) |
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and pseudo-max (100 - alpha percentile). Defaults to 1. |
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beta: absorbance threshold for transparent pixels. Defaults to 0.15 |
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max_cref: reference maximum stain concentrations for Hematoxylin & Eosin (H&E). |
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Defaults to (1.9705, 1.0308). |
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Note: |
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For more information refer to: |
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- the original paper: Macenko et al., 2009 http://wwwx.cs.unc.edu/~mn/sites/default/files/macenko2009.pdf |
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- the previous implementations: |
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- MATLAB: https://github.com/mitkovetta/staining-normalization |
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- Python: https://github.com/schaugf/HEnorm_python |
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""" |
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def __init__( |
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self, tli: float = 240, alpha: float = 1, beta: float = 0.15, max_cref: tuple | np.ndarray = (1.9705, 1.0308) |
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) -> None: |
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self.tli = tli |
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self.alpha = alpha |
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self.beta = beta |
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self.max_cref = np.array(max_cref) |
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def _deconvolution_extract_stain(self, image: np.ndarray) -> np.ndarray: |
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"""Perform Stain Deconvolution and return stain matrix for the image. |
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Args: |
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image: uint8 RGB image to perform stain deconvolution on |
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Return: |
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he: H&E absorbance matrix for the image (first column is H, second column is E, rows are RGB values) |
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""" |
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if not isinstance(image, np.ndarray): |
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raise TypeError("Image must be of type numpy.ndarray.") |
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if image.min() < 0: |
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raise ValueError("Image should not have negative values.") |
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if image.max() > 255: |
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raise ValueError("Image should not have values greater than 255.") |
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image = image.reshape((-1, 3)) |
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image = image.astype(np.float32, copy=False) + 1.0 |
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absorbance = -np.log(image.clip(max=self.tli) / self.tli) |
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absorbance_hat = absorbance[np.all(absorbance > self.beta, axis=1)] |
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if len(absorbance_hat) == 0: |
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raise ValueError("All pixels of the input image are below the absorbance threshold.") |
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_, eigvecs = np.linalg.eigh(np.cov(absorbance_hat.T).astype(np.float32, copy=False)) |
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t_hat = absorbance_hat.dot(eigvecs[:, 1:3]) |
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phi = np.arctan2(t_hat[:, 1], t_hat[:, 0]) |
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min_phi = np.percentile(phi, self.alpha) |
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max_phi = np.percentile(phi, 100 - self.alpha) |
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v_min = eigvecs[:, 1:3].dot(np.array([(np.cos(min_phi), np.sin(min_phi))], dtype=np.float32).T) |
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v_max = eigvecs[:, 1:3].dot(np.array([(np.cos(max_phi), np.sin(max_phi))], dtype=np.float32).T) |
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if v_min[0] > v_max[0]: |
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he = np.array((v_min[:, 0], v_max[:, 0]), dtype=np.float32).T |
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else: |
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he = np.array((v_max[:, 0], v_min[:, 0]), dtype=np.float32).T |
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return he |
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def __call__(self, image: np.ndarray) -> np.ndarray: |
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"""Perform stain extraction. |
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Args: |
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image: uint8 RGB image to extract stain from |
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return: |
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target_he: H&E absorbance matrix for the image (first column is H, second column is E, rows are RGB values) |
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""" |
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if not isinstance(image, np.ndarray): |
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raise TypeError("Image must be of type numpy.ndarray.") |
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target_he = self._deconvolution_extract_stain(image) |
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return target_he |
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class NormalizeHEStains(Transform): |
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"""Class to normalize patches/images to a reference or target image stain (see Note). |
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Performs stain deconvolution of the source image using the ExtractHEStains |
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class, to obtain the stain matrix and calculate the stain concentration matrix |
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for the image. Then, performs the inverse Beer-Lambert transform to recreate the |
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patch using the target H&E stain matrix provided. If no target stain provided, a default |
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reference stain is used. Similarly, if no maximum stain concentrations are provided, a |
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reference maximum stain concentrations matrix is used. |
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Args: |
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tli: transmitted light intensity. Defaults to 240. |
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alpha: tolerance in percentile for the pseudo-min (alpha percentile) and |
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pseudo-max (100 - alpha percentile). Defaults to 1. |
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beta: absorbance threshold for transparent pixels. Defaults to 0.15. |
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target_he: target stain matrix. Defaults to ((0.5626, 0.2159), (0.7201, 0.8012), (0.4062, 0.5581)). |
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max_cref: reference maximum stain concentrations for Hematoxylin & Eosin (H&E). |
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Defaults to [1.9705, 1.0308]. |
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Note: |
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For more information refer to: |
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- the original paper: Macenko et al., 2009 http://wwwx.cs.unc.edu/~mn/sites/default/files/macenko2009.pdf |
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- the previous implementations: |
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- MATLAB: https://github.com/mitkovetta/staining-normalization |
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- Python: https://github.com/schaugf/HEnorm_python |
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""" |
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def __init__( |
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self, |
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tli: float = 240, |
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alpha: float = 1, |
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beta: float = 0.15, |
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target_he: tuple | np.ndarray = ((0.5626, 0.2159), (0.7201, 0.8012), (0.4062, 0.5581)), |
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max_cref: tuple | np.ndarray = (1.9705, 1.0308), |
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) -> None: |
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self.tli = tli |
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self.target_he = np.array(target_he) |
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self.max_cref = np.array(max_cref) |
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self.stain_extractor = ExtractHEStains(tli=self.tli, alpha=alpha, beta=beta, max_cref=self.max_cref) |
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def __call__(self, image: np.ndarray) -> np.ndarray: |
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"""Perform stain normalization. |
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Args: |
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image: uint8 RGB image/patch to be stain normalized, pixel values between 0 and 255 |
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Return: |
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image_norm: stain normalized image/patch |
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""" |
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if not isinstance(image, np.ndarray): |
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raise TypeError("Image must be of type numpy.ndarray.") |
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if image.min() < 0: |
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raise ValueError("Image should not have negative values.") |
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if image.max() > 255: |
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raise ValueError("Image should not have values greater than 255.") |
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he = self.stain_extractor(image) |
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h, w, _ = image.shape |
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image = image.reshape((-1, 3)) |
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image = image.astype(np.float32) + 1.0 |
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absorbance = -np.log(image.clip(max=self.tli) / self.tli) |
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y = np.reshape(absorbance, (-1, 3)).T |
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conc = np.linalg.lstsq(he, y, rcond=None)[0] |
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max_conc = np.asarray([np.percentile(conc[0, :], 99), np.percentile(conc[1, :], 99)], dtype=np.float32) |
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tmp = np.divide(max_conc, self.max_cref, dtype=np.float32) |
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image_c = np.divide(conc, tmp[:, np.newaxis], dtype=np.float32) |
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image_norm: np.ndarray = np.multiply(self.tli, np.exp(-self.target_he.dot(image_c)), dtype=np.float32) |
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image_norm[image_norm > 255] = 254 |
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image_norm = np.reshape(image_norm.T, (h, w, 3)).astype(np.uint8) |
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return image_norm |
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