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Confocal Fluorescence Microscopy

Confocal Fluorescence Microscopy

Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...

Super-resolution Fluorescence Microscopy

Super-resolution Fluorescence Microscopy

Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been developed.

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Related Experiment Video

Updated: May 9, 2026

Confocal Imaging of Confined Quiescent and Flowing Colloid-polymer Mixtures

Confocal Imaging of Confined Quiescent and Flowing Colloid-polymer Mixtures

Published on: May 20, 2014

Simplified confocal microscope for counting particles at low concentrations.

Joseph P Skinner¹, Kerry M Swift, Qiaoqiao Ruan

¹Diagnostics Research, Abbott Diagnostics Division, Abbott Park, Illinois 60064, USA.

The Review of Scientific Instruments

|August 2, 2013

Summary

This summary is machine-generated.

This study presents a compact scanning confocal fluorescence microscope for rapid particle detection. The system achieves high sensitivity, detecting as few as 5 particles/ml and quantifying E. coli cells at 30 cells/ml.

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Last Updated: May 9, 2026

Confocal Imaging of Confined Quiescent and Flowing Colloid-polymer Mixtures

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Published on: May 20, 2014

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High-plex Imaging using Spectral Confocal Microscopy to Minimize Non-specific Tissue Fluorescence

High-plex Imaging using Spectral Confocal Microscopy to Minimize Non-specific Tissue Fluorescence

Published on: October 28, 2025

Area of Science:

Analytical Chemistry
Biotechnology
Microscopy

Background:

Accurate and rapid quantification of particles in liquid samples is crucial for various scientific and industrial applications.
Existing methods for particle detection can be time-consuming or lack the required sensitivity for low concentrations.

Purpose of the Study:

To develop and characterize a compact scanning confocal fluorescence microscope for sensitive particle detection.
To evaluate the system's performance in terms of detection limit, linearity, and statistical behavior.
To demonstrate the microscope's applicability for quantifying biological cells.

Main Methods:

A compact scanning confocal fluorescence microscope was designed, incorporating a mechanically moving cuvette and a large confocal volume.
Particle detection was achieved by analyzing spikes in fluorescence intensity data using pattern recognition as particles traversed the confocal volume.
The system was tested using fluorescent microspheres and stained biological cells (E. coli, yeast) at various concentrations.

Main Results:

The microscope demonstrated a detection sensitivity of 5 particles/ml for 3 μm fluorescent microspheres.
Linearity of detection was shown as a function of particle concentration.
The system successfully quantified E. coli cells at concentrations as low as 30 cells/ml within 10 minutes per sample.

Conclusions:

The developed compact scanning confocal fluorescence microscope offers a sensitive and rapid method for particle detection.
The system's ability to quantify low concentrations of biological cells highlights its potential for diverse applications.
This technology provides a valuable tool for high-throughput analysis in fields requiring precise particle enumeration.