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Confocal Fluorescence Microscopy

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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,...

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Video-rate Scanning Confocal Microscopy and Microendoscopy

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Portable low-cost and highly accurate programmable triggering controller for confocal spinning disk image scanning

Eli Flaxer¹, Alona Flaxer², Moran Sapir²

¹AFEKA - Tel-Aviv Academic College of Engineering, 69107 Tel-Aviv, Israel; School of Chemistry, The Sackler Faculty of Exact Sciences, Tel-Aviv University, 69978 Tel-Aviv, Israel.

Ultramicroscopy

|April 9, 2023

Summary

This summary is machine-generated.

This study introduces a low-cost digital signal controller (DSC) for confocal spinning-disk image scanning microscopy (CSD-ISM) triggering. This alternative achieves high accuracy and nanosecond resolution, making advanced microscopy more accessible.

Keywords:

Fluorescence microscopy Image scanning microscope Spinning-disk microscope

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Area of Science:

Microscopy and Imaging Technologies
Control Systems Engineering
Biophysics

Background:

Confocal spinning-disk image scanning microscopy (CSD-ISM) requires precise triggering for high-resolution imaging.
Previous triggering systems often rely on expensive field-programmable gate arrays (FPGAs), limiting accessibility.
There is a need for cost-effective and accurate triggering solutions in advanced microscopy.

Purpose of the Study:

To develop and validate a low-cost, high-accuracy triggering control system for CSD-ISM.
To demonstrate an alternative to expensive FPGA-based controllers.
To enhance the accessibility of advanced microscopy techniques.

Main Methods:

Implementation of a triggering controller using a low-cost real-time digital signal controller (DSC) development kit.
Connection of the DSC to a computer via an external USB channel.
Programming the DSC using free, included software, integrated with the Micro-Manager open-source microscopy platform.

Main Results:

Achieved a time resolution better than 10 nanoseconds for the triggering control.
Demonstrated a significantly lower cost compared to traditional FPGA-based systems.
Enabled flexible and virtually limitless output signal control through software calculations.

Conclusions:

The developed DSC-based triggering system offers a highly accurate and cost-effective solution for CSD-ISM.
This approach democratizes access to advanced microscopy by reducing hardware costs.
The system's performance and flexibility make it a viable alternative for various imaging applications.