Histogram-based analysis for confocal microscope images with immunofluorescent staining: A graphical-user-interface tool
Immunofluorescent staining is widely utilized in biomedical research. However, reliable and reproducible quantification remains challenging and often depend on the experience of the raters heavily. As a result, variations caused by different raters or the same rater at different times confound quant...
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| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-06-01
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| Series: | MethodsX |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215016125001529 |
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| Summary: | Immunofluorescent staining is widely utilized in biomedical research. However, reliable and reproducible quantification remains challenging and often depend on the experience of the raters heavily. As a result, variations caused by different raters or the same rater at different times confound quantitative interpretations. In this study, we propose a histogram-based analytical method to explore the sources of signals at various intensities and, subsequently, identify the signals of interest using the histograms as a reference. Our method aims to alleviate inter-rater and intra-rater inconsistencies and improve the quantification of microscope images with immunofluorescent staining. • Identification of signal sources for image pixels at different intensities • Reduction of quantification variations using the proposed histogram-based analysis |
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| ISSN: | 2215-0161 |