Simultaneous two-color imaging with a dual-channel miniscope in freely behaving mice

Miniaturized fluorescence microscopes (miniscopes) enable imaging of calcium events from a large population of neurons in freely behaving animals. Traditionally, miniscopes have only been able to record from a single fluorescence wavelength. Here, we present an open-source dual-channel miniscope tha...

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Published inScience advances Vol. 11; no. 27; p. eadr6470
Main Authors Dong, Zhe, Feng, Yu, Diego, Keziah, Baggetta, Austin M., Sweis, Brian M., Pennington, Zachary T., Lamsifer, Sophia I., Zaki, Yosif, Sangiuliano, Federico, Philipsberg, Paul A., Morales-Rodriguez, Denisse, Kircher, Daniel, Slesinger, Paul, Shuman, Tristan, Aharoni, Daniel, Cai, Denise J.
Format Journal Article
LanguageEnglish
Published United States American Association for the Advancement of Science 04.07.2025
Subjects
Animals
Behavior, Animal
CA1 Region, Hippocampal - diagnostic imaging
CA1 Region, Hippocampal - metabolism
Calcium - metabolism
Cellular Neuroscience
Hippocampus
Mice
Microscopy, Fluorescence - instrumentation
Microscopy, Fluorescence - methods
Neurons - metabolism
Neuroscience
SciAdv r-articles
Online AccessGet full text
ISSN2375-2548
2375-2548
DOI10.1126/sciadv.adr6470

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Abstract Miniaturized fluorescence microscopes (miniscopes) enable imaging of calcium events from a large population of neurons in freely behaving animals. Traditionally, miniscopes have only been able to record from a single fluorescence wavelength. Here, we present an open-source dual-channel miniscope that simultaneously records two wavelengths in freely behaving animals. To enable simultaneous acquisition of two fluorescent wavelengths, we incorporated two CMOS sensors into a single miniscope. To validate our dual-channel miniscope, we imaged hippocampal CA1 region that co-expressed a dynamic calcium indicator (GCaMP) and a static nuclear signal (dTomato) while mice ran on a linear track. Our results suggest that, even when neurons were registered across days using dTomato signals, hippocampal spatial coding changes over time. In conclusion, our dual-channel miniscope enables imaging of two fluorescence wavelengths with minimal cross-talk between the two channels, opening the doors to a multitude of previously inaccessible experimental possibilities. An open-source dual-channel miniscope that simultaneously records two wavelengths in freely behaving animals was presented.
AbstractList Miniaturized fluorescence microscopes (miniscopes) enable imaging of calcium events from a large population of neurons in freely behaving animals. Traditionally, miniscopes have only been able to record from a single fluorescence wavelength. Here, we present an open-source dual-channel miniscope that simultaneously records two wavelengths in freely behaving animals. To enable simultaneous acquisition of two fluorescent wavelengths, we incorporated two CMOS sensors into a single miniscope. To validate our dual-channel miniscope, we imaged hippocampal CA1 region that co-expressed a dynamic calcium indicator (GCaMP) and a static nuclear signal (dTomato) while mice ran on a linear track. Our results suggest that, even when neurons were registered across days using dTomato signals, hippocampal spatial coding changes over time. In conclusion, our dual-channel miniscope enables imaging of two fluorescence wavelengths with minimal cross-talk between the two channels, opening the doors to a multitude of previously inaccessible experimental possibilities. An open-source dual-channel miniscope that simultaneously records two wavelengths in freely behaving animals was presented.
Miniaturized fluorescence microscopes (miniscopes) enable imaging of calcium events from a large population of neurons in freely behaving animals. Traditionally, miniscopes have only been able to record from a single fluorescence wavelength. Here, we present an open-source dual-channel miniscope that simultaneously records two wavelengths in freely behaving animals. To enable simultaneous acquisition of two fluorescent wavelengths, we incorporated two CMOS sensors into a single miniscope. To validate our dual-channel miniscope, we imaged hippocampal CA1 region that co-expressed a dynamic calcium indicator (GCaMP) and a static nuclear signal (dTomato) while mice ran on a linear track. Our results suggest that, even when neurons were registered across days using dTomato signals, hippocampal spatial coding changes over time. In conclusion, our dual-channel miniscope enables imaging of two fluorescence wavelengths with minimal cross-talk between the two channels, opening the doors to a multitude of previously inaccessible experimental possibilities.Miniaturized fluorescence microscopes (miniscopes) enable imaging of calcium events from a large population of neurons in freely behaving animals. Traditionally, miniscopes have only been able to record from a single fluorescence wavelength. Here, we present an open-source dual-channel miniscope that simultaneously records two wavelengths in freely behaving animals. To enable simultaneous acquisition of two fluorescent wavelengths, we incorporated two CMOS sensors into a single miniscope. To validate our dual-channel miniscope, we imaged hippocampal CA1 region that co-expressed a dynamic calcium indicator (GCaMP) and a static nuclear signal (dTomato) while mice ran on a linear track. Our results suggest that, even when neurons were registered across days using dTomato signals, hippocampal spatial coding changes over time. In conclusion, our dual-channel miniscope enables imaging of two fluorescence wavelengths with minimal cross-talk between the two channels, opening the doors to a multitude of previously inaccessible experimental possibilities.
Miniaturized fluorescence microscopes (miniscopes) enable imaging of calcium events from a large population of neurons in freely behaving animals. Traditionally, miniscopes have only been able to record from a single fluorescence wavelength. Here, we present an open-source dual-channel miniscope that simultaneously records two wavelengths in freely behaving animals. To enable simultaneous acquisition of two fluorescent wavelengths, we incorporated two CMOS sensors into a single miniscope. To validate our dual-channel miniscope, we imaged hippocampal CA1 region that co-expressed a dynamic calcium indicator (GCaMP) and a static nuclear signal (dTomato) while mice ran on a linear track. Our results suggest that, even when neurons were registered across days using dTomato signals, hippocampal spatial coding changes over time. In conclusion, our dual-channel miniscope enables imaging of two fluorescence wavelengths with minimal cross-talk between the two channels, opening the doors to a multitude of previously inaccessible experimental possibilities.
Author Baggetta, Austin M.
Dong, Zhe
Sweis, Brian M.
Philipsberg, Paul A.
Aharoni, Daniel
Kircher, Daniel
Diego, Keziah
Pennington, Zachary T.
Morales-Rodriguez, Denisse
Feng, Yu
Lamsifer, Sophia I.
Slesinger, Paul
Shuman, Tristan
Zaki, Yosif
Sangiuliano, Federico
Cai, Denise J.
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Snippet Miniaturized fluorescence microscopes (miniscopes) enable imaging of calcium events from a large population of neurons in freely behaving animals....
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StartPage eadr6470
SubjectTerms Animals
Behavior, Animal
CA1 Region, Hippocampal - diagnostic imaging
CA1 Region, Hippocampal - metabolism
Calcium - metabolism
Cellular Neuroscience
Hippocampus
Mice
Microscopy, Fluorescence - instrumentation
Microscopy, Fluorescence - methods
Neurons - metabolism
Neuroscience
SciAdv r-articles
Title Simultaneous two-color imaging with a dual-channel miniscope in freely behaving mice
URI https://www.ncbi.nlm.nih.gov/pubmed/40601747
https://www.proquest.com/docview/3226713665
https://pubmed.ncbi.nlm.nih.gov/PMC12219470
Volume 11
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