Journal
/
Cancer Research
/
Industrialized, Artificial Intelligence-guided Laser Microdissection for Microscaled Proteomic Analysis of the Tumor Microenvironment
JoVE Journal
Cancer Research
This content is Free Access.
JoVE Journal Cancer Research
Industrialized, Artificial Intelligence-guided Laser Microdissection for Microscaled Proteomic Analysis of the Tumor Microenvironment

Industrialized, Artificial Intelligence-guided Laser Microdissection for Microscaled Proteomic Analysis of the Tumor Microenvironment

DOI:
10.3791/64171-v

•

13:01 min

•

June 03, 2022

•

, , , , , , , ,
1Women’s Health Integrated Research Center, Gynecologic Cancer Center of Excellence, Department of Obstetrics and Gynecology,Uniformed Services University and Walter Reed National Military Medical Center, 2The Henry M. Jackson Foundation for the Advancement of Military Medicine, 3The John P. Murtha Cancer Center Research Program, Department of Surgery,Uniformed Services University, 4Women’s Health Integrated Research Center, Inova Women’s Service Line,Inova Health System

Chapters

  • 00:04Introduction
  • 01:07LMD Slide(s) Preparation
  • 01:58Automated Feature Selection Using Image Analysis Software
  • 06:58Laser Microdissectionvvv
  • 09:41Protein Digestion by Pressure-Cycle Technology (PCT) and Sample Analysis Using Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)
  • 10:55Results: Analysis of the Proteins in LMD-Enriched and Whole Tumor Samples
  • 12:09Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol describes a high-throughput workflow for artificial intelligence-driven segmentation of pathology-confirmed regions of interest from stained, thin tissue section images for enrichment of histology-resolved cell populations using laser microdissection. This strategy includes a novel algorithm enabling the transfer of demarcations denoting cell populations of interest directly to laser microscopes.

Tags

AI-driven Image AnalysisLaser MicrodissectionProteomic AnalysisTumor MicroenvironmentPathomicsCalibration FiducialsPEN MembraneImage Analysis SoftwareAnnotationsPartitioningTiled AnnotationsPython Dapa Algorithm

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/64171?article_type=v

A Mimic of the Tumor Microenvironment: A Simple Method for Generating Enriched Cell Populations and Investigating Intercellular Communication
check_url/64171?article_type=v

Conditional Knockdown of Gene Expression in Cancer Cell Lines to Study the Recruitment of Monocytes/Macrophages to the Tumor Microenvironment
check_url/64171?article_type=v

Simple and Rapid Method to Obtain High-quality Tumor DNA from Clinical-pathological Specimens Using Touch Imprint Cytology
check_url/64171?article_type=v

Digital Polymerase Chain Reaction Assay for the Genetic Variation in a Sporadic Familial Adenomatous Polyposis Patient Using the Chip-in-a-tube Format
check_url/64171?article_type=v

Measuring Bone Remodeling and Recreating the Tumor-Bone Microenvironment Using Calvaria Co-culture and Histomorphometry
check_url/64171?article_type=v

Assessing Tumor Microenvironment of Metastasis Doorway-Mediated Vascular Permeability Associated with Cancer Cell Dissemination using Intravital Imaging and Fixed Tissue Analysis
check_url/64171?article_type=v

Analysis of Side Population in Solid Tumor Cell Lines
check_url/64171?article_type=v

Isolation of Proximal Fluids to Investigate the Tumor Microenvironment of Pancreatic Adenocarcinoma
check_url/64171?article_type=v

Generation of Zebrafish Larval Xenografts and Tumor Behavior Analysis
check_url/64171?article_type=v

A Label-Free Segmentation Approach for Intravital Imaging of Mammary Tumor Microenvironment

Read Article