Background:Color consistency in histology images is still an issue in digital pathology. Different imaging systems reproduced the colors of a histological slide differently.Materials and Methods:Color correction was implemented using the color information of the nine color patches of a color calibration slide. The inherent spectral colors of these patches along with their scanned colors were used to derive a color correction matrix whose coefficients were used to convert the pixels’ colors to their target colors.Results:There was a significant reduction in the CIELAB color difference, between images of the same H & E histological slide produced by two different whole slide scanners by 3.42 units, P < 0.001 at 95% confidence level.Conclusion:Color variations in histological images brought about by whole slide scanning can be effectively normalized with the use of the color calibration slide.

Color enhancement of multispectral images is useful to visualize the image's spectral features. Previously, a color enhancement method, which enhances the feature of a specified spectral band without changing the average color distribution, was proposed. However, sometimes the enhanced features are indiscernible or invisible, especially when the enhanced spectrum lies outside the visible range. In this paper, we extended the conventional method for more effective visualization of the spectral features both in visible range and non-visible range. In the proposed method, the user specifies both the spectral band for extracting the spectral feature and the color for visualization respectively, so that the spectral feature is enhanced with arbitrary color. The proposed color enhancement method was applied to different types of multispectral images where its effectiveness to visualize spectral features was verified.

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Objectives: 3D histology tissue modeling is a useful analytical technique for understanding anatomy and disease at the cellular level. However, the current accuracy of 3D histology technology is largely unknown, and errors, misalignment and missing information are common in 3D tissue reconstruction. We used micro-CT imaging technology to better understand these issues and the relationship between fresh tissue and its 3D histology counterpart. Methods: We imaged formalin-fixed and 2% Lugol-stained mouse brain, human uterus and human lung tissue with micro-CT. We then conducted image analyses on the tissues before and after paraffin embedding using 3D Slicer and ImageJ software to understand how tissue changes between the fixation and embedding steps. Results: We found that all tissue samples decreased in volume by 19.2-61.5% after embedding, that micro-CT imaging can be used to assess the integrity of tissue blocks, and that micro-CT analysis can help to design an optimized tissue-sectioning protocol. Conclusions: Micro-CT reference data help to identify where and to what extent tissue was lost or damaged during slide production, provides valuable anatomical information for reconstructing missing parts of a 3D tissue model, and aids in correcting reconstruction errors when fitting the image information in vivo and ex vivo.

Objective:The image quality in whole slide imaging (WSI) is one of the most important issues for the practical use of WSI scanners. In this paper, we proposed an image quality evaluation method for scanned slide images in which no reference image is required.Methods:While most of the conventional methods for no-reference evaluation only deal with one image degradation at a time, the proposed method is capable of assessing both blur and noise by using an evaluation index which is calculated using the sharpness and noise information of the images in a given training data set by linear regression analysis. The linear regression coefficients can be determined in two ways depending on the purpose of the evaluation. For objective quality evaluation, the coefficients are determined using a reference image with mean square error as the objective value in the analysis. On the other hand, for subjective quality evaluation, the subjective scores given by human observers are used as the objective values in the analysis. The predictive linear regression models for the objective and subjective image quality evaluations, which were constructed using training images, were then used on test data wherein the calculated objective values are construed as the evaluation indices.Results:The results of our experiments confirmed the effectiveness of the proposed image quality evaluation method in both objective and subjective image quality measurements. Finally, we demonstrated the application of the proposed evaluation method to the WSI image quality assessment and automatic rescanning in the WSI scanner.