Multiphoton Microscopy of FITC-labelled Fusobacterium nucleatum in a Mouse in vivo Model of Breast Cancer

Lishay Parhi*, Amjad Shhadeh, Naseem Maalouf, Tamar Alon-Maimon, Viviana Scaiewicz, Gilad Bachrach

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Over the past decades, the main techniques used to visualize bacteria in tissue have improved but are still mainly based on indirect recognition of bacteria. Both microscopy and molecular recognition are being improved, but most procedures for bacteria detection in tissue involve extensive damage. Here, we describe a method to visualize bacteria in tissue slices from an in vivo model of breast cancer. This method allows examining trafficking and colonization of fluorescein-5-isothiocyanate (FITC)-stained bacteria in various tissues. The protocol provides direct visualization of fusobacterial colonization in breast cancer tissue. Rather than processing the tissue or confirming bacterial colonization by PCR or culture, the tissue is directly imaged using multiphoton microscopy. This direct visualization protocol causes no damage to the tissue; therefore, all structures can be identified. This method can be combined with others to co-visualize bacteria, types of cells, or protein expression in cells.

Original languageAmerican English
Article numbere4635
JournalBio-protocol
Volume13
Issue number6
DOIs
StatePublished - 20 Mar 2023

Bibliographical note

Publisher Copyright:
Copyright: © 2023 The Authors; exclusive licensee Bio-protocol LLC.

Keywords

  • Bacterial colonization
  • Breast cancer
  • Fusobacterium nucleatum
  • Multiphoton microscopy

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