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Integrated nanoplasmonic sensing for cellular functional immunoanalysis using human blood.

Bo-Ram Oh1, Nien-Tsu Huang, Weiqiang Chen

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Summary
This summary is machine-generated.

This study introduces a novel localized surface plasmon resonance (LSPR) biosensing technique for detecting tumor necrosis factor-alpha (TNF-α) secreted by immune cells. The method significantly reduces sample volume and assay time for cytokine detection in human blood.

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Area of Science:

  • Nanotechnology and Bionanosensing
  • Immunology and Immunoassays
  • Optofluidics and Microfluidics

Background:

  • Localized surface plasmon resonance (LSPR) offers label-free, real-time biomolecule detection but faces challenges in sensitivity and sample handling for cytokine secretion assays.
  • Cytokine secretion assays are crucial for assessing immune cell function and donor immune status, yet conventional methods are often time-consuming and require large sample volumes.
  • Previous applications of LSPR bionanosensing have not addressed cytokine detection from clinically relevant immune cells in human blood.

Purpose of the Study:

  • To develop and validate a label-free LSPR biosensing technique for detecting tumor necrosis factor-alpha (TNF-α) secreted by immune cells in clinical blood samples.
  • To integrate LSPR bionanosensors with an optofluidic platform for enhanced sensitivity and reduced sample handling in cellular functional immunoanalysis.

Main Methods:

  • Development of an optofluidic platform integrating LSPR bionanosensors for trapping and stimulating immune cells within a microfluidic chamber.
  • Utilizing on-chip spatial confinement to increase local cytokine concentration for rapid detection by the LSPR setup.
  • Testing the developed biosensing technique with THP-1 cells and subsequently with CD45 cells isolated directly from human blood.

Main Results:

  • The LSPR optofluidics device successfully detected TNF-α secreted by as few as 1000 cells, achieving a ~100-fold reduction in required sample volume compared to conventional assays.
  • The assay demonstrated cellular functional immunoanalysis using a minimal blood sample volume of 3 μL.
  • The total assay time was reduced by three times compared to conventional enzyme-linked immunosorbent assay (ELISA) methods.

Conclusions:

  • The developed label-free LSPR optofluidic biosensing technique enables sensitive and rapid detection of cytokines from minimal blood samples.
  • This approach overcomes key limitations of previous LSPR bionanosensing methods for cytokine secretion assays, offering a significant advancement in cellular functional immunoanalysis.
  • The technique holds promise for efficient assessment of immune status and cellular immune responses using clinically relevant samples.