Home › Microscopy & Imaging › Optical Screening of Novel Bacteria-specific Probes on Ex Vivo Human Lung Tissue by Confocal Laser Endomicroscopy
Microscopy & Imaging JoVE (Open Access) Citable · DOI

Optical Screening of Novel Bacteria-specific Probes on Ex Vivo Human Lung Tissue by Confocal Laser Endomicroscopy

DOI: 10.3791/56284-v
What you'll learn
  • ✓ Prepare and handle ex vivo human lung tissue for optical imaging
  • ✓ Configure and operate confocal laser endomicroscopy (CLE) device
  • ✓ Collect and analyze fluorescence microscopy data from bacterial probes
  • ✓ Evaluate bacteria-specific optical imaging agents in tissue samples
Protocol

This technique describes an efficient screening process for evaluating bacteria-specific optical imaging agents within ex vivo human lung tissue, by fibered confocal fluorescence microscopy for the rapid identification of small molecule chemical probe-candidates with translatable potential.

Difficulty
advanced
Total time
~3–4 hours per tissue sample (including device setup, imaging acquisition, and analysis)
Biosafety
BSL-2

Steps

1
Prepare ex vivo human lung tissue

Obtain and prepare human lung tissue samples for imaging, ensuring tissue viability and appropriate handling for downstream confocal microscopy analysis.

▶ 00:44
2
Setup confocal laser endomicroscopy device

Calibrate and configure the CLE device, including laser alignment, detection parameters, and optical path optimization for high-resolution bacterial imaging.

▶ 01:53
3
Collect fluorescence data using CLE

Acquire confocal images of bacteria-specific optical probes within lung tissue, capturing z-stack and multi-wavelength fluorescence data.

▶ 03:55
4
Analyze microscopy data and probe performance

Process and quantify fluorescence signals, evaluate probe-bacteria colocalization, and assess candidate probes for translational potential.

▶ 05:59
5
Interpret FCFM imaging results in tissue context

Review representative images showing bacterial labeling patterns within human lung tissue architecture and determine probe efficacy.

▶ 07:41
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