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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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Related Experiment Video

Updated: Dec 29, 2025

Detection of Fluorescent Nanoparticle Interactions with Primary Immune Cell Subpopulations by Flow Cytometry

Detection of Fluorescent Nanoparticle Interactions with Primary Immune Cell Subpopulations by Flow Cytometry

Published on: March 28, 2014

Labelling primary immune cells using bright blue fluorescent nanoparticles.

Maura C Belanger¹, Meng Zhuang², Alexander G Ball³

¹Department of Chemistry, University of Virginia, Charlottesville, Virginia 22904, USA. rrp2z@virginia.edu fraser@virginia.edu and Beirne B. Carter Center for Immunology Research, University of Virginia, Charlottesville, Virginia 22903, USA.

Biomaterials Science

|February 7, 2020

Summary

This summary is machine-generated.

New boron-based fluorescent nanoparticles offer a bright, UV-excitable option for short-term cell tracking. These difluoroboron β-diketonate nanoparticles effectively label cells without impacting viability or immune response, improving imaging in live tissue.

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Area of Science:

Biomaterials Science
Cell Biology
Fluorescence Imaging

Background:

Cell tracking is vital for biological studies, requiring reagents that don't alter cell state or autofluorescence.
Existing UV-excitable reagents are limited, despite advances in violet-excited dyes.
Difluoroboron β-diketonates are known UV-excitable bio-imaging agents with high quantum yields.

Purpose of the Study:

To investigate difluoroboron dibenzoylmethane (BF2dbm) dye-polymer conjugates as potential cell tracking reagents.
To synthesize and characterize nanoparticles for UV-based cell imaging.
To evaluate the efficacy and biocompatibility of these nanoparticles for cell labeling.

Main Methods:

Synthesis of BF2dbm conjugates with biocompatible polymers (PLLA, PCL, PEG).
Fabrication of dye-polymer conjugates into nanoparticles.
Assessment of nanoparticle stability, cell labeling efficiency, cellular viability, and immune response.
Comparison with a commercial UV-excitable cytoplasmic labeling reagent in live tissue imaging.

Main Results:

Nanoparticles were stable in water and cell culture media for one week but aggregated in saline.
Phagocytic splenocytes were more effectively labeled by the nanoparticles.
Cellular viability and basic immune responses were unaffected by nanoparticle labeling.
Labeled cells were more distinguishable in live tissue compared to commercial reagents.

Conclusions:

BF2dbm-polymer nanoparticles are a promising short-term cell labeling strategy.
Their high fluorescence and labeling efficiency enhance imaging in biological contexts.
Wide excitation peaks offer versatility across different imaging platforms.
These nanoparticles address the need for effective UV-excitable cell tracking reagents.