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Conducting Multiple Imaging Modes with One Fluorescence Microscope

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

Wedge prism approach for simultaneous multichannel microscopy.

Hanna Cai¹, Yao L Wang², Richard T Wainner³

¹GAMDAN Optics Inc., 2362 Qume Drive, Suite B, San Jose, CA, 95131, USA. hanna.cai@gamdan.com.

Scientific Reports

|November 30, 2019

Summary

This summary is machine-generated.

Researchers developed a novel, inexpensive add-on for multichannel optical microscopy using wedge prisms. This simple device enables simultaneous multicolor imaging for biological research, offering a user-friendly alternative to complex commercial systems.

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Area of Science:

Optical Microscopy
Biophotonics
Neuroscience

Background:

Multichannel (multicolor) imaging is crucial for various biological applications, including in vivo neuronal calcium imaging and fluorescent label colocalization.
Existing commercial instruments often rely on complex and expensive filter-switching or dual-view strategies, posing accessibility challenges.

Purpose of the Study:

To introduce a novel, simple, and cost-effective add-on device for simultaneous multichannel optical microscopy.
To demonstrate the efficacy of wedge prisms for multicolor imaging applications in biological research.

Main Methods:

Developed a novel add-on module utilizing simple wedge prisms for simultaneous multichannel optical microscopy.
Performed point spread function measurements and Zemax simulations to evaluate optical performance and resolution effects.
Tested a two-channel device on live C. elegans neurons and a four-channel device on fixed chick embryo Brainbow samples.

Main Results:

The wedge prism device requires no alignment, is robust, user-friendly, and significantly less expensive than current commercial alternatives.
Optical simulations indicated a reduction in resolution orthogonal to the wedge interface and axially, without introducing aberrations, with effects most pronounced at the field of view periphery.
Experimental results showed comparable imaging signals to conventional dual-view instruments in C. elegans and enabled spectral identification of neurons in Brainbow samples without extensive postprocessing.

Conclusions:

The wedge prism-based multichannel imaging approach offers a practical and affordable solution for simultaneous multicolor microscopy.
This technology has the potential for widespread adoption in diverse microscopy applications within biological research.
The device provides a valuable alternative for researchers seeking efficient and cost-effective multicolor imaging capabilities.