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Quantitative histological image analyses of reticulin fibers in a myelofibrotic mouse
Hector A Lucero1, Shenia Patterson1, Shinobu Matsuura1
1Departments of Medicine and Biochemistry, Whitaker Cardiovascular Institute, Boston University School of Medicine, Boston, MA 02118, USA.
Abstract:
Bone marrow (BM) reticulin fibrosis (RF), revealed by silver staining of tissue sections, is associated with myeloproliferative neoplasms, while tools for quantitative assessment of reticulin deposition throughout a femur BM are still in need. Here, we present such a method, allowing via analysis of hundreds of composite images to identify a patchy nature of RF throughout the BM during disease progression in a mouse model of myelofibrosis. To this end, initial conversion of silver stained BM color images into binary images identified two limitations: variable color, owing to polychromatic staining of reticulin fibers, and variable background in different sections of the same batch, limiting application of the color deconvolution method, and use of constant threshold, respectively. By blind coding image identities, to allow for threshold input (still within a narrow range), and using shape filtering to further eliminate background we were able to quantitate RF in myelofibrotic Gata-1low (experimental) and wild type (control) mice as a function of animal age. Color images spanning the whole femur BM were batch-analyzed using ImageJ software, aided by our two newly added macros. The results show heterogeneous RF density in different areas of the marrow of Gata-1low mice, with degrees of heterogeneity reduced upon aging. This method can be applied uniformly across laboratories in studies assessing RF remodeling induced by aging or other conditions in animal models.
Insights
A new method quantifies bone marrow reticulin fibrosis (RF) in mouse models. This technique reveals patchy RF distribution and heterogeneity, aiding research into myeloproliferative neoplasms and myelofibrosis.
Area of Science:
- Biomedical Engineering
- Hematology
- Pathology
Background:
- Reticulin fibrosis (RF) in bone marrow (BM) is linked to myeloproliferative neoplasms.
- Quantitative assessment tools for BM reticulin deposition are lacking.
- Accurate quantification is crucial for understanding disease progression.
Purpose of the Study:
- To develop a quantitative method for assessing reticulin fibrosis in whole femur bone marrow.
- To analyze the spatial distribution and progression of RF in a mouse model of myelofibrosis.
- To overcome limitations of existing methods for reticulin staining analysis.
Main Methods:
- Developed a quantitative method using ImageJ software and custom macros for analyzing silver-stained femur BM images.
- Converted color images to binary images and applied shape filtering to address variable staining and background.
- Quantified RF in Gata-1low (myelofibrotic) and wild-type mice across different ages.
Main Results:
- Identified a patchy, heterogeneous distribution of reticulin fibrosis in the bone marrow of myelofibrotic mice.
- Observed a reduction in RF heterogeneity with aging in the Gata-1low mouse model.
- Demonstrated the method's ability to quantitate RF across the entire femur BM.
Conclusions:
- The developed method enables quantitative assessment of reticulin fibrosis in bone marrow.
- The findings highlight the heterogeneous nature of RF and its modulation by aging.
- This technique is applicable across laboratories for studying RF remodeling in various conditions and animal models.
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