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Updated: May 6, 2026

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Published on: June 30, 2018

An open-source alignment method for multichannel infinite-conjugate microscopes using a ray transfer matrix analysis

Gemma S Cairns¹, Brian R Patton

¹Department of Physics and SUPA, University of Strathclyde , Glasgow G4 0NG, UK.

Philosophical Transactions. Series A, Mathematical, Physical, and Engineering Sciences

|June 3, 2024

Summary

This summary is machine-generated.

This study introduces a new algorithm for aligning microscope lenses and cameras using only image magnification. This method simplifies alignment for open-source microscopes, achieving accuracy comparable to the microscope's axial resolution.

Keywords:

open-source microscopy optical alignment ray transfer matrix analysis

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Area of Science:

Optical Engineering
Microscopy
Open-Source Hardware

Background:

Multichannel optical systems enable multiple imaging modes in a single microscope.
Traditional alignment methods require extra hardware, posing challenges for compact, open-source designs.

Purpose of the Study:

To develop a simplified alignment algorithm for multichannel microscopes.
To enable precise lens and camera positioning using only image magnification measurements.
To validate the algorithm's effectiveness in open-source microscope systems.

Main Methods:

Developed an alignment algorithm based on image magnification measurements.
Utilized ray transfer matrix analysis to model optical paths and magnifications.
Tested the algorithm on M4All and OpenFlexure open-source microscope platforms.

Main Results:

Achieved positioning accuracy comparable to the microscope's axial resolution.
Successfully determined objective lens misalignment (working distance) and required adjustments.
Validated the algorithm's usability and effectiveness in real-world open-source microscope applications.

Conclusions:

The proposed image-based alignment method simplifies optical system setup.
This technique is suitable for compact, open-source microscope designs.
The open-source implementation facilitates reproducible microscopy hardware development.