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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

Depiction of the lipofuscin quantitation. Three images of the same section, where (a) is the autofluorescence image in the red area, (b) is the quantitated lipofuscin, and (c) is a combination of (a and b) with (b) made slightly translucent and layered on top of (a) for overview reasons
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Figure 8: Depiction of the lipofuscin quantitation. Three images of the same section, where (a) is the autofluorescence image in the red area, (b) is the quantitated lipofuscin, and (c) is a combination of (a and b) with (b) made slightly translucent and layered on top of (a) for overview reasons

Mentions: As for verification of the lipofuscin quantitation, no comparison can be done to the VG-stained sections. Figure 8 shows an example of lipofuscin content analysis. Panel a is an autofluorescence image in the red area. The outline mask of the corresponding digitally quantitated lipofuscin is shown in Panel b. Panel c is panel a with a slightly translucent Panel b on top.


Quantitative analysis of myocardial tissue with digital autofluorescence microscopy.

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

Depiction of the lipofuscin quantitation. Three images of the same section, where (a) is the autofluorescence image in the red area, (b) is the quantitated lipofuscin, and (c) is a combination of (a and b) with (b) made slightly translucent and layered on top of (a) for overview reasons
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 8: Depiction of the lipofuscin quantitation. Three images of the same section, where (a) is the autofluorescence image in the red area, (b) is the quantitated lipofuscin, and (c) is a combination of (a and b) with (b) made slightly translucent and layered on top of (a) for overview reasons
Mentions: As for verification of the lipofuscin quantitation, no comparison can be done to the VG-stained sections. Figure 8 shows an example of lipofuscin content analysis. Panel a is an autofluorescence image in the red area. The outline mask of the corresponding digitally quantitated lipofuscin is shown in Panel b. Panel c is panel a with a slightly translucent Panel b on top.

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