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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

Scatterplots of point grid counting versus digital quantitations of cells (a), extracellular phase (b), and fibrous tissue (c). X = Y is added as a red dashed line, black full line is least squares fitted line, and the green lines display the 95% confidence intervals thereof
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Figure 5: Scatterplots of point grid counting versus digital quantitations of cells (a), extracellular phase (b), and fibrous tissue (c). X = Y is added as a red dashed line, black full line is least squares fitted line, and the green lines display the 95% confidence intervals thereof

Mentions: Figure 4a shows an example of a point grid quantitated area along with corresponding digital quantitation of ECP [Figure 4b], fibrous tissue [Figure 4c], and lipofuscin [Figure 4d]. Fibrous tissue, ECP, and cells were quantified in the point grid analysis. It was not possible to quantify lipofuscin using the VG-stain because the lipofuscin could not be differentiated from surrounding myocyte cytosol. Scatter plots displaying the correlations between point grid quantitations and digital quantitations are presented in Figure 5. The digital quantitation of cells is the entire area of the section with fibrous tissue and ECP subtracted. An evident correlation is noted. The function y = x is entirely included in confidence intervals of the cell and ECP quantitations. The confidence interval of the fibrous tissue quantitation is slightly below x = y in the central part of the fit corresponding to a small systematic underestimation of fibrous tissue in the digital quantitation.


Quantitative analysis of myocardial tissue with digital autofluorescence microscopy.

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

Scatterplots of point grid counting versus digital quantitations of cells (a), extracellular phase (b), and fibrous tissue (c). X = Y is added as a red dashed line, black full line is least squares fitted line, and the green lines display the 95% confidence intervals thereof
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 5: Scatterplots of point grid counting versus digital quantitations of cells (a), extracellular phase (b), and fibrous tissue (c). X = Y is added as a red dashed line, black full line is least squares fitted line, and the green lines display the 95% confidence intervals thereof
Mentions: Figure 4a shows an example of a point grid quantitated area along with corresponding digital quantitation of ECP [Figure 4b], fibrous tissue [Figure 4c], and lipofuscin [Figure 4d]. Fibrous tissue, ECP, and cells were quantified in the point grid analysis. It was not possible to quantify lipofuscin using the VG-stain because the lipofuscin could not be differentiated from surrounding myocyte cytosol. Scatter plots displaying the correlations between point grid quantitations and digital quantitations are presented in Figure 5. The digital quantitation of cells is the entire area of the section with fibrous tissue and ECP subtracted. An evident correlation is noted. The function y = x is entirely included in confidence intervals of the cell and ECP quantitations. The confidence interval of the fibrous tissue quantitation is slightly below x = y in the central part of the fit corresponding to a small systematic underestimation of fibrous tissue in the digital quantitation.

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