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Related Concept Videos

UV–Vis Spectrometers01:14

UV–Vis Spectrometers

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The absorbance of UV and visible (UV–visible) radiations is measured using a UV–visible spectrophotometer. Deuterium lamps, which emit UV radiation, and tungsten lamps, which produce radiation in the visible region, are used as light sources in UV–visible spectrophotometers. A monochromator or prism is used for diffraction grating, i.e., to split the incoming radiation into different wavelengths. A system of slits is used to focus the desired wavelength on the sample cell.
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Related Experiment Video

Updated: Jun 17, 2025

Diffuse Reflectance Spectroscopy: Getting the Capillary Refill Test Under One's Thumb
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Published on: December 2, 2017

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Senescence detection using reflected light.

Benjamin Dedic1, Leo Westerberg1, Andrea Mosqueda Solís1,2

  • 1Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm, Sweden.

Aging Cell
|August 5, 2024
PubMed
Summary
This summary is machine-generated.

We developed a novel reflected light method to accurately quantify senescent cells, particularly in human adipocytes. This technique improves detection sensitivity for senescence-associated beta-galactosidase (SABG) activity, aiding research into obesity and Type 2 diabetes.

Keywords:
SABGX‐galadipocytesbeta‐galactosidasebrightfieldreflected lightsenescencewestern blot

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Area of Science:

  • Cellular Biology
  • Aging Research
  • Metabolic Disease

Background:

  • Cellular senescence plays roles in development, tissue repair, cancer, and aging.
  • Increased senescence is linked to obesity and Type 2 diabetes (T2D).
  • Quantifying senescent cells, especially in large primary cells like human adipocytes, is challenging.

Purpose of the Study:

  • To present a novel reflected light-based method for accurate senescence-associated beta-galactosidase (SABG) staining measurements.
  • To enable precise quantification of senescent cells in various cell types, including primary human adipocytes.
  • To offer a superior alternative to traditional methods for studying adipocyte senescence and its link to metabolic disorders.

Main Methods:

  • Utilized reflected light microscopy with confocal imaging to detect X-gal crystals in SABG-stained cells.
  • Integrated reflected light measurements with immunofluorescence for comprehensive cell state classification.
  • Incorporated sample-specific pH controls for robust senescence assessment.
  • Validated the technique in primary human and mouse adipocytes, and differentiated 3T3-L1 cells.

Main Results:

  • Achieved superior sensitivity in detecting X-gal precipitates compared to traditional brightfield techniques.
  • Captured diverse X-gal staining patterns, revealing improved detection sensitivity.
  • Demonstrated that reflected light analysis provides a more accurate quantification of SABG activity in mature human adipocytes than western blot.
  • Showcased the method's compatibility with immunofluorescence and various cell types.

Conclusions:

  • The novel reflected light approach offers a highly sensitive and accurate method for quantifying cellular senescence, particularly in adipocytes.
  • This technique facilitates a deeper understanding of adipocyte senescence and its implications in obesity and Type 2 diabetes.
  • Reflected light microscopy provides a more comprehensive and sensitive tool for senescence research than conventional methods.