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Related Experiment Videos

High-throughput Screening of Erratic Cell Volume Regulation Using a Hydrogel-based Single-cell Microwell Array.

Casey L Brown1, Valerie Fleischauer, Jinseok Heo

  • 1The State University of New York College at Buffalo.

Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry
|April 11, 2017
PubMed
Summary

A novel microwell array system enables high-throughput screening of cells with defective regulatory volume decrease (RVD). This method efficiently identifies cells lacking normal RVD, crucial for maintaining cell volume homeostasis.

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Area of Science:

  • Cell biology
  • Biophysics
  • Biotechnology

Background:

  • Regulatory volume decrease (RVD) is essential for cell volume homeostasis in hypotonic environments.
  • Understanding RVD mechanisms is critical, but high-throughput screening of individual cell responses has been challenging.

Purpose of the Study:

  • To develop and validate a high-throughput single-cell screening method for defective RVD.
  • To quantify the prevalence of impaired RVD in Madin-Darby canine kidney (MDCK) cells.

Main Methods:

  • Utilized a photocrosslinked hydrogel microwell array for single-cell culture.
  • Employed a volume-sensitive fluorescent dye and wide-field fluorescence microscopy to monitor cell volume changes.
  • Analyzed volume dynamics of over 100 single cells simultaneously using time-lapse imaging in hypotonic conditions.

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Main Results:

  • Successfully screened single Madin-Darby canine kidney (MDCK) cells for defective RVD.
  • Identified that approximately 40% of MDCK cells exhibited weak or absent RVD.
  • Demonstrated superior capacity for analyzing discrete single-cell volume changes compared to previous methods.

Conclusions:

  • The developed microwell array platform provides an efficient high-throughput method for RVD screening.
  • The findings highlight a significant proportion of MDCK cells with impaired RVD, suggesting complex regulatory mechanisms.
  • This approach is expected to accelerate the elucidation of RVD mechanisms in various cell types.