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Quantitative analysis of myocardial tissue with digital autofluorescence microscopy.

Jensen T, Holten-Rossing H, Svendsen IM, Jacobsen C, Vainer B - J Pathol Inform (2016)

Bottom Line: This data may provide a basic histological starting point from which further digital analysis including staining may benefit.The presented method is amply described as a prestain multicomponent quantitation and outlining tool for histological sections of cardiac tissue.The main perspective is the opportunity for combination with digital analysis of stained microsections, for which the method may provide an accurate digital framework.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Rigshospitalet, University of Copenhagen, Copenhagen, Denmark.

ABSTRACT

Background: The opportunity offered by whole slide scanners of automated histological analysis implies an ever increasing importance of digital pathology. To go beyond the importance of conventional pathology, however, digital pathology may need a basic histological starting point similar to that of hematoxylin and eosin staining in conventional pathology. This study presents an automated fluorescence-based microscopy approach providing highly detailed morphological data from unstained microsections. This data may provide a basic histological starting point from which further digital analysis including staining may benefit.

Methods: This study explores the inherent tissue fluorescence, also known as autofluorescence, as a mean to quantitate cardiac tissue components in histological microsections. Data acquisition using a commercially available whole slide scanner and an image-based quantitation algorithm are presented.

Results: It is shown that the autofluorescence intensity of unstained microsections at two different wavelengths is a suitable starting point for automated digital analysis of myocytes, fibrous tissue, lipofuscin, and the extracellular compartment. The output of the method is absolute quantitation along with accurate outlines of above-mentioned components. The digital quantitations are verified by comparison to point grid quantitations performed on the microsections after Van Gieson staining.

Conclusion: The presented method is amply described as a prestain multicomponent quantitation and outlining tool for histological sections of cardiac tissue. The main perspective is the opportunity for combination with digital analysis of stained microsections, for which the method may provide an accurate digital framework.

No MeSH data available.


Related in: MedlinePlus

Examples of fluorescence images. (a and b) The same area in the blue and the red filter, respectively. (c and d) The corresponding ratio images with (c) being the ratio of blue to red and (d) the ratio of red to blue. Intensity histograms with fitted standard deviations corresponding to the myocyte peaks are shown in the small insets. Van Gieson-staining and quantitations of the central parts of this microsection are displayed in Figure 2
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Figure 1: Examples of fluorescence images. (a and b) The same area in the blue and the red filter, respectively. (c and d) The corresponding ratio images with (c) being the ratio of blue to red and (d) the ratio of red to blue. Intensity histograms with fitted standard deviations corresponding to the myocyte peaks are shown in the small insets. Van Gieson-staining and quantitations of the central parts of this microsection are displayed in Figure 2

Mentions: The obtained autofluorescence images were in focus with level contrast and intensity. All images were considered suitable for further analysis. Examples are given in Figure 1. Particular differences between the two wavelengths are noted. In the blue filter [Figure 1a], the intensity of fibrous tissue was high and the intensity of lipofuscin was low. The opposite holds true for the red filter [Figure 1b]. The images can be regarded as different stainings simultaneously of one microsection. No bleaching artifacts from autofocus could be identified. The inserted intensity histograms include fitted standard deviations corresponding to the myocytes. In both wavelengths, they were considered normally distributed.


Quantitative analysis of myocardial tissue with digital autofluorescence microscopy.

Jensen T, Holten-Rossing H, Svendsen IM, Jacobsen C, Vainer B - J Pathol Inform (2016)

Examples of fluorescence images. (a and b) The same area in the blue and the red filter, respectively. (c and d) The corresponding ratio images with (c) being the ratio of blue to red and (d) the ratio of red to blue. Intensity histograms with fitted standard deviations corresponding to the myocyte peaks are shown in the small insets. Van Gieson-staining and quantitations of the central parts of this microsection are displayed in Figure 2
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4837794&req=5

Figure 1: Examples of fluorescence images. (a and b) The same area in the blue and the red filter, respectively. (c and d) The corresponding ratio images with (c) being the ratio of blue to red and (d) the ratio of red to blue. Intensity histograms with fitted standard deviations corresponding to the myocyte peaks are shown in the small insets. Van Gieson-staining and quantitations of the central parts of this microsection are displayed in Figure 2
Mentions: The obtained autofluorescence images were in focus with level contrast and intensity. All images were considered suitable for further analysis. Examples are given in Figure 1. Particular differences between the two wavelengths are noted. In the blue filter [Figure 1a], the intensity of fibrous tissue was high and the intensity of lipofuscin was low. The opposite holds true for the red filter [Figure 1b]. The images can be regarded as different stainings simultaneously of one microsection. No bleaching artifacts from autofocus could be identified. The inserted intensity histograms include fitted standard deviations corresponding to the myocytes. In both wavelengths, they were considered normally distributed.

Bottom Line: This data may provide a basic histological starting point from which further digital analysis including staining may benefit.The presented method is amply described as a prestain multicomponent quantitation and outlining tool for histological sections of cardiac tissue.The main perspective is the opportunity for combination with digital analysis of stained microsections, for which the method may provide an accurate digital framework.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Rigshospitalet, University of Copenhagen, Copenhagen, Denmark.

ABSTRACT

Background: The opportunity offered by whole slide scanners of automated histological analysis implies an ever increasing importance of digital pathology. To go beyond the importance of conventional pathology, however, digital pathology may need a basic histological starting point similar to that of hematoxylin and eosin staining in conventional pathology. This study presents an automated fluorescence-based microscopy approach providing highly detailed morphological data from unstained microsections. This data may provide a basic histological starting point from which further digital analysis including staining may benefit.

Methods: This study explores the inherent tissue fluorescence, also known as autofluorescence, as a mean to quantitate cardiac tissue components in histological microsections. Data acquisition using a commercially available whole slide scanner and an image-based quantitation algorithm are presented.

Results: It is shown that the autofluorescence intensity of unstained microsections at two different wavelengths is a suitable starting point for automated digital analysis of myocytes, fibrous tissue, lipofuscin, and the extracellular compartment. The output of the method is absolute quantitation along with accurate outlines of above-mentioned components. The digital quantitations are verified by comparison to point grid quantitations performed on the microsections after Van Gieson staining.

Conclusion: The presented method is amply described as a prestain multicomponent quantitation and outlining tool for histological sections of cardiac tissue. The main perspective is the opportunity for combination with digital analysis of stained microsections, for which the method may provide an accurate digital framework.

No MeSH data available.


Related in: MedlinePlus