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Quantification of Confocal Images Using LabVIEW for Tissue Engineering Applications.

Lauren Sfakis1, Tim Kamaldinov1, Melinda Larsen2

  • 11 SUNY Polytechnic Institute , Nanobioscience Constellation, Albany, New York.

Tissue Engineering. Part C, Methods
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Summary

A new LabVIEW program, Bio-LIME, quantifies 3D protein localization in confocal images for tissue engineering. It aids in evaluating scaffold efficacy and understanding cell polarity crucial for tissue regeneration.

Keywords:
LabVIEWimage processingnanofiberssalivary gland developmenttissue engineering

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Area of Science:

  • Biomedical Engineering
  • Cell Biology
  • Image Analysis

Background:

  • Accurate quantification of protein localization in 3D is critical for tissue engineering (TE) applications, particularly for evaluating scaffold constructs and cell differentiation.
  • Understanding protein positioning within cells grown on scaffolds is key to assessing scaffold efficacy for tissue regeneration.
  • Functional epithelial cells require specific apicobasal polarity, with proteins correctly localized to apical or basolateral regions.

Purpose of the Study:

  • To develop a customized program for quantifying protein positions in 3D confocal image Z-stacks of epithelial cell monolayers.
  • To demonstrate the utility of this program in salivary gland tissue engineering and assess protein localization changes in response to scaffold modifications.
  • To evaluate the impact of nanofiber scaffold dimensions on epithelial apicobasal polarization.

Main Methods:

  • Development of a LabVIEW-based program named Bio-LabVIEW Image Matrix Evaluation (Bio-LIME).
  • Bio-LIME processes 3D confocal Z-stack images to determine cell and nuclei dimensions, protein localization, and cell counts.
  • Application of Bio-LIME to quantify the 3D location of Zonula occludens-1 protein and assess epithelial polarization on different scaffolds.

Main Results:

  • Bio-LIME successfully quantifies 3D protein localization, cell height, nuclei dimensions, and cell count from confocal Z-stack images.
  • The program demonstrated changes in Zonula occludens-1 protein 3D position after scaffold chemical modification with laminin.
  • No advantage was found for sub-100 nm poly lactic-co-glycolic acid nanofibers over 250 nm fibers regarding epithelial apicobasal polarization.

Conclusions:

  • Bio-LIME provides a robust method for quantifying protein localization in 3D, essential for tissue engineering and regeneration research.
  • The developed program aids in evaluating scaffold efficacy by analyzing protein positioning and cell organization.
  • Bio-LIME has broad applicability for 3D protein quantification in various biological contexts, facilitating advancements in cell biology and tissue engineering.