.png){kind=icon}
返回
Senescence detection using reflected light
.png){kind=icon}
OA
AI
DOI：10.1111/acel.14295.png)
.png){kind=icon}
.png){kind=icon}
分享.png){kind=icon}
收藏
摘要
En 中文
Senescence is an important cellular program occurring in development, tissue repair, cancer, and aging. Increased senescence is also associated with disease states, including obesity and Type 2 diabetes (T2D). Characterizing and quantifying senescent cells at a single cell level has been challenging and particularly difficult in large primary cells, such as human adipocytes. In this study, we present a novel approach that utilizes reflected light for accurate senescence-associated beta-galactosidase (SABG) staining measurements, which can be integrated with immunofluorescence and is compatible with primary mature adipocytes from both human and mouse, as well as with differentiated 3T3-L1 cells. This technique provides a more comprehensive classification of a cell's senescent state by incorporating multiple criteria, including robust sample-specific pH controls. By leveraging the precision of confocal microscopy to detect X-gal crystals using reflected light, we achieved superior sensitivity over traditional brightfield techniques. This strategy allows for the capture of all X-gal precipitates in SABG-stained samples, revealing diverse X-gal staining patterns and improved detection sensitivity. Additionally, we demonstrate that reflected light outperforms western blot analysis for the detection and quantification of senescence in mature human adipocytes, as it offers a more accurate representation of SABG activity. This detection strategy enables a more thorough investigation of senescent cell characteristics and specifically a deeper look at the relationship between adipocyte senescence and obesity associated disorders, such as T2D. Senescence-associated beta-galactosidase (SABG) activity was measured using reflected light microscopy by replacing the dichroic mirror with a T80/R20 mirror. SABG activity was manipulated by adjusting the pH, providing both a positive control and a threshold for SABG positivity, enabling objective quantification of senescence. This method is compatible with simultaneous antibody staining.image
Keyword:
adipocytes
beta-galactosidase
brightfield
reflected light
SABG
senescence
western blot
X-gal
AI总结
对已上传原文的论文进行重点信息的提取，主要内容包括：简要概述、研究摘要、背景介绍、关键亮点、图文解析、展望与总结。
期刊
IF:
7.1
论文数:
3.8K
被引数:
2.3W
机构
引用论文
.png){kind=icon}
分享收藏In vitro model of chronological aging of adipocytes: Interrelationships with hypoxia and oxidation
PREAI
分享收藏
查看更多内容