Nonlinear optical microscopy: use of second harmonic generation and two-photon microscopy for automated quantitative liver fibrosis studies (original) (raw)
Related papers
2000
Background/Aims: Liver fibrosis is one of the most important and characteristic histologic alterations in progressive and chronic liver diseases. Thus, in both clinical and experimental practice, it is fundamental to have a reliable and objective method for its precise quantification. Several semi-quantitative scoring systems have been described. All are time-consuming and produce partially subjective fibrosis evaluations that are not very precise. This paper describes the design and validation of an original image analysis-based application, FibroQuant, for automatically and rapidly quantifying perisinusoidal, perivenular and portalperiportal and septal fibrosis and portal-periportal and septal morphology in liver histologic specimens. Methods: The implemented image-processing algorithms automatically segment interstitial fibrosis areas, while extraction of portal-periportal and septal region is carried out with an automatic algorithm and a simple interactive step. For validation, all automatically extracted areas were also manually segmented and quantified.
Automated Whole Slide Image Analysis for a Translational Quantification of Liver Fibrosis
Current literature highlights the need for precise histological quantitative assessment of fibrosis which cannot be achieved by conventional scoring systems, inherent to their discontinuous values and reader-dependent variability. Here we used an automated image analysis software to quantitate fibrosis deposition in two relevant preclinical models of liver fibrosis, and established correlation with other quantitative fibrosis descriptors. Longitudinal quantification of liver fibrosis was carried out during progression of post-necrotic (CCl4-induced) and metabolic (HF-CDAA feeding) models of chronic liver disease in mice. Whole slide images of picrosirius red-stained liver sections were analyzed in fully automated, unsupervised manner by our software. Fibrosis was characterized by a significant increase of collagen proportionate area (CPA) at weeks 3 (CCl4) and 8 (HF-CDAA) with a progressive increase up to week 18 and 24, respectively. CPA was compared to collagen content assessed bi...
Assessment of Hepatic Fibrosis by Image Analysis Software in relation to Physiological Markers
Background: The chronic liver disease is one of the most deadly diseases in Egypt and around the world; perfect diagnosis methods are urgently needed. The proper assessment of liver fibrosis offers acute information for both perfect diagnosis and accurate therapeutic decision-making. Although digital image analysis (DIA) is a promising method for quantitative assessment of liver fibrosis, it has not been fully evaluated in practice yet. The present study was designed to explore the possibilities of using open access image analysis software (ImageJ) for accurate quantitative assessment of liver fibrosis. Methods: This study investigated the use of digital images analysis by software ImageJ of liver sections stained by sirius red (SR). Reproducibility of this technique was tested in comparison with the a semi-quantitative score of liver fibrosis as well as other markers of fibrosis including physiological markers of liver functions such as alanine transaminase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT). In addition, it was tested compared to serum marker collagen IV (Col IV) and hepatic hydroxyproline. Results: From physiological point of view; hepatic hydroxyproline, serum Col IV and ALT showed higher accuracy, sensitivity and specificity than other physiological markers with area under curve (AUC) of 0.935, 0.955 and 0.902 respectively. On the other hand, DIA of SR by ImageJ software had shown the best accuracy with AUC of 1.0. In conclusions: our novel data prove that the use of ImageJ with semiautomatic color segmentation is a reliable and practical way of measuring collagen proportionate area.
A robust collagen scoring method for human liver fibrosis by second harmonic microscopy
Optics Express, 2010
Second Harmonic Generation (SHG) microscopy offers the opportunity to image collagen of type I without staining. We recently showed that a simple scoring method, based on SHG images of histological human liver biopsies, correlates well with the Metavir assessment of fibrosis level (Gailhouste et al., J. Hepatol., 2010). In this article, we present a detailed study of this new scoring method with two different objective lenses. By using measurements of the objectives point spread functions and of the photomultiplier gain, and a simple model of the SHG intensity, we show that our scoring method, applied to human liver biopsies, is robust to the objective's numerical aperture (NA) for low NA, the choice of the reference sample and laser power, and the spatial sampling rate. The simplicity and robustness of our collagen scoring method may open new opportunities in the quantification of collagen content in different organs, which is of main importance in providing diagnostic information and evaluation of therapeutic efficiency. Stone, "Urinary assays for desmosine and hydroxylysylpyridinoline in the detection of cirrhosis,"
Scandinavian journal of gastroenterology, 2017
Liver fibrosis is traditionally graded into categorical stages with cirrhosis as the highest stage. However, cirrhosis stage may vary between individuals widely in terms of the amount of fibrosis which is not assessed by traditional staging systems. We aimed to utilise visual morphometry to quantify the amount of fibrosis in liver biopsy and compare how non-invasive methods of quantifying liver fibrosis correlated with histological measures. Liver biopsy specimens from 115 consecutive chronic liver disease patients were assessed by a single pathologist for fibrosis stage by the Clinical Research Network and METAVIR systems as well as percentage fibrosis by visual morphometry. Liver T1 relaxation times, liver stiffness measurement (LSM) by transient elastography and enhanced liver fibrosis (ELF) score were compared between fibrosis stages. In addition, these parameters were correlated with pathologist's visual estimate of percentage fibrosis and their predictive ability for advan...