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Influence of light and mounting medium on the fading of Feulgen stain
Abstract:
Serial microspectrophotometric estimations of the absorbance of Feulgen-stained interphase nuclei of human fibroblasts were made on slides mounted in a variety of mounting media and stored in light or in darkness. All preparations showed some fading, at rates which varied with different mountants, were greater in the light than in the dark, and were greater in the monochromatic illumination of the microspectrophotometer under working conditions than in normal laboratory light conditions. Fading was minimised by using XAM improved white neutral mounting medium (G.T. Gurr), storing slides in darkenss, and by reading samples as quickly as possible.
Insights
Feulgen-stained nuclei absorbance fades over time, especially in light. Storing slides in darkness and using specific mounting media like XAM minimizes this DNA staining fading.
Area of Science:
- Cell Biology
- Histology
- Spectrophotometry
Background:
- Feulgen staining is a cytological technique used to stain DNA.
- Microspectrophotometry is a technique used to measure absorbance of light by microscopic samples.
- Staining intensity can fade over time, affecting quantitative analysis.
Purpose of the Study:
- To investigate the fading rates of Feulgen-stained human fibroblast nuclei.
- To evaluate the impact of different mounting media and storage conditions on staining stability.
- To identify methods for minimizing Feulgen stain fading.
Main Methods:
- Serial microspectrophotometric absorbance measurements of Feulgen-stained interphase nuclei.
- Comparison of various mounting media.
- Evaluation of storage in light versus darkness.
- Assessment of fading under microspectrophotometer illumination.
Main Results:
- All preparations exhibited some degree of Feulgen stain fading.
- Fading rates varied significantly depending on the mounting medium used.
- Staining faded more rapidly when stored in light compared to darkness.
- Fading was accelerated under the monochromatic light of the microspectrophotometer.
Conclusions:
- Mounting medium choice, light exposure, and measurement duration significantly influence Feulgen stain stability.
- XAM improved white neutral mounting medium, dark storage, and rapid sample analysis minimize fading.
- Optimized protocols are crucial for reliable quantitative microspectrophotometry of Feulgen-stained nuclei.