Method for optimising rapid staining to visualise structural changes in mammalian sperm chromatin
The optimized sperm chromatin visualization method addresses time and reproducibility issues by using modified agarose, denaturing solutions, and staining techniques, enhancing image quality and consistency for routine analysis.
WO2026139646A1PCT designated stage Publication Date: 2026-07-02CHROMACELL SL
Patent Information
- Authority / Receiving Office
- WO · WO
- Patent Type
- Applications
- Current Assignee / Owner
- CHROMACELL SL
- Filing Date
- 2024-12-23
- Publication Date
- 2026-07-02
AI Technical Summary
Technical Problem
Existing methods for visualizing structural changes in sperm chromatin are time-consuming, lack reproducibility, and provide inconsistent staining results, especially in cases of poor semen quality, and are not suitable for routine clinical analysis.
Method used
A modified procedure involving optimized use of low-melting-point agarose microgel, reduced denaturing and deproteinizing solutions, cold ethanol fixation, and panoptic staining to enhance sperm chromatin visualization in bright-field and fluorescence microscopy.
Benefits of technology
The method significantly reduces processing time, improves image quality, and provides consistent differentiation of sperm DNA fragmentation, suitable for fresh, frozen, or cryopreserved samples.
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Smart Images
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Abstract
The invention relates to an improved method for analysing sperm DNA structure, by means of which it is possible to reduce the time required to process samples by 80%, optimising every stage of the process. This effect is achieved through a synergetic improvement in each step of the process. The formation of microgels is accelerated with cold metal plates (2–8°C) and using 0.8%-1.5% agarose. DNA denaturation is carried out using HCl (100 mM). The deproteinisation solution contains DTT (0.22 M), NaCl (2.75 M), detergents of the alcohol ethoxylate (0.044 M) and SDS (0.0014 M) type. Fixation is carried out using 96% ethanol at a low temperature. The staining process uses a panoptic dye, improving the quality of the images. The invention not only speeds up the analysis by reducing the time required by 80%, but also enables faster and more accurate diagnoses, thanks to the high quality of the images obtained.
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