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Lossless Immunocytochemistry Using Photo-Polymerized Hydrogel Thin-Films

Journal
Analyst, 2020, 145(8), 2897–2903

Overview

This work addresses a recurring problem in immunocytochemistry workflows:
cells get lost during staining and wash cycles — especially when cell number is low, cells are fragile, or do not naturally adhere to substrates.

We developed a photo-polymerized PEG hydrogel thin-film that encapsulates cells during staining.
The film:

  • stabilizes cells during reagent exchange
  • remains optically clear for imaging
  • allows antibody diffusion
  • requires no specialized equipment beyond a UV source

My Role

  • Developed and optimized the hydrogel thin-film polymerization workflow (gel formulation, curing conditions, thickness control)
  • Performed experimental validation showing that hydrogel encapsulation prevents cell loss across repeated wash cycles
  • Evaluated antibody penetration and fluorescence intensity consistency across encapsulated vs non-encapsulated controls
  • Imaging, quantitative analysis, and interpretation of single-cell retention and signal uniformity
  • Assisted in preparing figures and manuscript revisions, particularly workflow visualization and comparative retention metrics

Key Contributions

  • Demonstrated nearly lossless retention of suspension and low-adherence cell types during staining
  • Verified compatibility with common immunocytochemistry reagents, fluorophores, and microscopy modalities
  • Provided a simple, low-cost workflow suitable for research and clinical imaging pipelines

Why It Matters

Traditional immobilization methods (surface coatings, microfluidics, centrifugation) introduce:

  • variable adhesion
  • cellular stress
  • positional drift
  • or require specialized equipment

This hydrogel-based immobilization maintains biological context while keeping workflows accessible.

It supports future applications in:

  • diagnostic imaging
  • single-cell analysis pipelines
  • imaging-driven cell selection and downstream omics workflows

➡️ Read more publications → /publications/

Vaughn Lee