Home › Microscopy & Imaging › Automated Slide Scanning and Segmentation in Fluorescently-labeled Tissues Using a Widefield High-content Analysis System
Microscopy & Imaging JoVE (Open Access) Citable · DOI

Automated Slide Scanning and Segmentation in Fluorescently-labeled Tissues Using a Widefield High-content Analysis System

DOI: 10.3791/57440-v
What you'll learn
  • ✓ Configure and execute automated preview scans on fluorescently-labeled tissue slides
  • ✓ Set up high-magnification imaging parameters for widefield high-content analysis
  • ✓ Perform automated segmentation and quantitation of fluorescent markers in tissues
  • ✓ Interpret representative images and analyze results from high-content scanning systems
Protocol

Here we describe a protocol for the automated segmentation of fluorescently labeled tissues on slides using a widefield high-content analysis system (WHCAS). This protocol has wide-ranging applications in any field which involves the quantitation of fluorescent markers in biological tissues including the biological sciences, medical engineering, and health sciences.

Difficulty
intermediate
Total time
~2–4 hours per slide set (including preview scan, setup, acquisition, and analysis)

Steps

1
Perform preview scan and configure magnification setup

Execute an initial preview scan of the fluorescently-labeled tissue slide and configure the widefield high-content analysis system for optimal high-magnification imaging parameters.

▶ 00:51
2
Acquire high-magnification images of tissue regions

Using the configured settings, perform automated high-magnification image acquisition across selected tissue regions on the slide to capture fluorescent marker distribution.

▶ 02:51
3
Segment and analyze fluorescent marker quantitation

Apply automated segmentation algorithms to the acquired images to quantify fluorescent markers within tissue regions, generating representative images and quantitative results.

▶ 07:54
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