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A Rapid Method for Multispectral Fluorescence Imaging of Frozen Tissue Sections
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UltraPlex Hapten-Based Multiplexed Fluorescent Immunohistochemistry.

Matt Levin1, Amy C Flor2, Helen Snyder1

  • 1Cell IDx, San Diego, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|July 31, 2021
PubMed
Summary

The UltraPlex method enables rapid, sensitive multiplexed fluorescent immunohistochemistry for imaging four or more antigens simultaneously in tissue samples. This technique enhances visualization of complex cellular subtypes, including tumor cells and lymphocytes.

Keywords:
AntibodyHaptenImmunofluorescenceImmunohistochemistryMicroscopyMultiplex imaging

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Area of Science:

  • Biomedical research
  • Immunohistochemistry
  • Cellular imaging

Background:

  • Multiplexed immunohistochemistry allows simultaneous detection of multiple antigens.
  • Existing methods can be complex and time-consuming.
  • High-resolution imaging of cellular subtypes is crucial for disease research.

Purpose of the Study:

  • To describe the UltraPlex method for multiplexed two-dimensional fluorescent immunohistochemistry.
  • To enable rapid and sensitive imaging of four or more antigens per tissue section.
  • To facilitate high-resolution visualization of complex cellular subtypes.

Main Methods:

  • Utilizes hapten tags conjugated to primary antibodies.
  • Employs fluorophore-labeled anti-hapten secondary antibodies for signal amplification.
  • Detection is performed using standard fluorescent microscopes or digital slide scanners.

Main Results:

  • Achieves multiplexed imaging of four or more antigens per tissue section.
  • Provides highly sensitive and specific staining.
  • Enables high-resolution imaging of multiple cellular subtypes, including tumor cells and tumor-infiltrating lymphocytes.
  • Digital overlay of 4-plex stained samples creates 8-plex (or more) high-content images.

Conclusions:

  • The UltraPlex method is a rapid, straightforward protocol for multiplexed immunohistochemistry.
  • It enables sensitive and specific detection of multiple antigens in conventionally prepared tissue samples.
  • Facilitates advanced visualization of cellular subtypes and their distribution in tissues.