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Immunofluorescence Microscopy

Immunofluorescence Microscopy

A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
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Establishing Fungal Entomopathogens as Endophytes: Towards Endophytic Biological Control

Establishing Fungal Entomopathogens as Endophytes: Towards Endophytic Biological Control

Published on: April 11, 2013

Efficient fluorescence-based localization technique for real-time tracking endophytes route in host-plants

Christine A Ondzighi-Assoume¹, Bandana Bhusal¹, Adam M Traore¹

¹College of Agriculture, Department of Agricultural and Environmental Sciences Tennessee State University Nashville Tennessee USA.

|August 12, 2022

Summary

This study developed a fluorescence-based method to track beneficial bacteria (BCAs) in plants. The technique successfully visualized bacterial colonization in roots and hypocotyls, aiding in understanding plant-microbe interactions.

Keywords:

Arabidopsis thaliana biological control agents colonization competency sweet sorghum tagged‐fluorescent bacteria transformation

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Monitoring Spatial Segregation in Surface Colonizing Microbial Populations

Monitoring Spatial Segregation in Surface Colonizing Microbial Populations

Published on: October 29, 2016

Area of Science:

Agricultural Microbiology
Plant Pathology
Molecular Biology

Background:

Biological control agents (BCAs) are crucial for sustainable agriculture, reducing reliance on chemical pesticides.
Effective tracking of BCAs within host plants is vital for understanding their efficacy and optimizing application strategies.

Purpose of the Study:

To develop a reliable fluorescence-based technique for labeling and tracking bacterial biological control agents (BCAs) in host-plant interactions.
To elucidate the localization and colonization routes of BCAs within plants.
To establish a reproducible method for assessing BCA movement and niches.

Main Methods:

Transformation of bacterial isolates (PSL, IMC8, PS) with plasmids carrying reporter genes (eGFP, pporRFP).
Development of antibiotic-resistant competent BCAs.
Fluorescence microscopy and PCR analysis to confirm BCA localization in Arabidopsis and sorghum.

Main Results:

High plasmid-mediated transformation efficiencies (up to 84%) were achieved for selected BCAs.
Fluorescently tagged BCAs were localized to plant roots and hypocotyls, but not leaves or stems.
Successful confirmation of BCA colonization using fluorescence imaging and PCR.

Conclusions:

The developed fluorescence-based technique provides high-resolution, reproducible tracking of BCAs within plant tissues.
This method enables spatial assessment of BCA movement and niches, advancing our understanding of host-plant interactions.
The protocol facilitates the study of biological and physiological mechanisms underlying plant-microbe interactions.