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Related Experiment Video

Updated: Sep 13, 2025

Multiplexed Barcoding Image Analysis for Immunoprofiling and Spatial Mapping Characterization in the Single-Cell Analysis of Paraffin Tissue Samples

Multiplexed Barcoding Image Analysis for Immunoprofiling and Spatial Mapping Characterization in the Single-Cell Analysis of Paraffin Tissue Samples

Published on: April 7, 2023

Protocol for processing and analyzing multiplexed images improves lymphatic cell identification and spatial

Justin A Smith¹, Drew T Bloss¹, MacKenzie D Williams¹

¹Department of Pathology, Immunology, and Laboratory Medicine, College of Medicine, Diabetes Institute, University of Florida, Gainesville, FL 32610, USA.

|July 27, 2025

Summary

We developed KINTSUGI, a user-guided image processing protocol for human lymphatic tissue. This method ensures quality control for cell phenotyping and spatial analysis, streamlining complex data preparation.

Keywords:

Computer sciences Immunology Microscopy Protein Biochemistry Proteomics

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Last Updated: Sep 13, 2025

Multiplexed Barcoding Image Analysis for Immunoprofiling and Spatial Mapping Characterization in the Single-Cell Analysis of Paraffin Tissue Samples

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Published on: September 22, 2013

Area of Science:

Immunology
Bioinformatics
Microscopy

Background:

Multiplexed imaging of human lymphatic tissue requires extensive preprocessing for accurate cell analysis.
Existing protocols can be complex and lack interactive quality control measures.

Purpose of the Study:

To introduce KINTSUGI, a novel, user-guided image processing protocol.
To ensure robust quality control throughout the analysis pipeline for multiplexed lymphatic tissue images.

Main Methods:

KINTSUGI protocol encompasses parameter tuning and batch processing.
Includes steps like illumination correction, stitching, deconvolution, 3D-2D conversion, registration, and autofluorescence subtraction.
Details segmentation, feature extraction, phenotyping, and spatial analysis procedures.

Main Results:

KINTSUGI provides interactive user engagement for quality control.
Streamlines preprocessing of raw image data for cell phenotyping and spatial analysis.
Facilitates detailed downstream analysis of lymphatic tissue architecture and cell populations.

Conclusions:

KINTSUGI offers a simplified, user-friendly approach to multiplexed image processing.
Enhances the reliability and reproducibility of cell phenotyping and spatial analysis in lymphatic research.
A valuable tool for researchers working with complex biological imaging data.