Live Cell Light Sheet Imaging with Low- and High-Spatial-Coherence Detection Approaches Reveals Spatiotemporal Aspects of Neuronal Signaling
Light sheet microscopy in live cells requires minimal excitation intensity and resolves three-dimensional (3D) information rapidly. Lattice light sheet microscopy (LLSM) works similarly but uses a lattice configuration of Bessel beams to generate a flatter, diffraction-limited z-axis sheet suitable...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-06-01
|
| Series: | Journal of Imaging |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2313-433X/9/6/121 |
| _version_ | 1797594068160610304 |
|---|---|
| author | Mariana Potcoava Donatella Contini Zachary Zurawski Spencer Huynh Christopher Mann Jonathan Art Simon Alford |
| author_facet | Mariana Potcoava Donatella Contini Zachary Zurawski Spencer Huynh Christopher Mann Jonathan Art Simon Alford |
| author_sort | Mariana Potcoava |
| collection | DOAJ |
| description | Light sheet microscopy in live cells requires minimal excitation intensity and resolves three-dimensional (3D) information rapidly. Lattice light sheet microscopy (LLSM) works similarly but uses a lattice configuration of Bessel beams to generate a flatter, diffraction-limited z-axis sheet suitable for investigating subcellular compartments, with better tissue penetration. We developed a LLSM method for investigating cellular properties of tissue in situ. Neural structures provide an important target. Neurons are complex 3D structures, and signaling between cells and subcellular structures requires high resolution imaging. We developed an LLSM configuration based on the Janelia Research Campus design or in situ recording that allows simultaneous electrophysiological recording. We give examples of using LLSM to assess synaptic function in situ. In presynapses, evoked Ca<sup>2+</sup> entry causes vesicle fusion and neurotransmitter release. We demonstrate the use of LLSM to measure stimulus-evoked localized presynaptic Ca<sup>2+</sup> entry and track synaptic vesicle recycling. We also demonstrate the resolution of postsynaptic Ca<sup>2+</sup> signaling in single synapses. A challenge in 3D imaging is the need to move the emission objective to maintain focus. We have developed an incoherent holographic lattice light-sheet (IHLLS) technique to replace the LLS tube lens with a dual diffractive lens to obtain 3D images of spatially incoherent light diffracted from an object as incoherent holograms. The 3D structure is reproduced within the scanned volume without moving the emission objective. This eliminates mechanical artifacts and improves temporal resolution. We focus on LLS and IHLLS applications and data obtained in neuroscience and emphasize increases in temporal and spatial resolution using these approaches. |
| first_indexed | 2024-03-11T02:17:28Z |
| format | Article |
| id | doaj.art-7b648747bfb64bde95eb5d18f9a18617 |
| institution | Directory Open Access Journal |
| issn | 2313-433X |
| language | English |
| last_indexed | 2024-03-11T02:17:28Z |
| publishDate | 2023-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Journal of Imaging |
| spelling | doaj.art-7b648747bfb64bde95eb5d18f9a186172023-11-18T11:04:30ZengMDPI AGJournal of Imaging2313-433X2023-06-019612110.3390/jimaging9060121Live Cell Light Sheet Imaging with Low- and High-Spatial-Coherence Detection Approaches Reveals Spatiotemporal Aspects of Neuronal SignalingMariana Potcoava0Donatella Contini1Zachary Zurawski2Spencer Huynh3Christopher Mann4Jonathan Art5Simon Alford6Department of Anatomy and Cell Biology, University of Illinois at Chicago, 808 South Wood Street, Rm 578 MC 512, Chicago, IL 60612, USADepartment of Anatomy and Cell Biology, University of Illinois at Chicago, 808 South Wood Street, Rm 578 MC 512, Chicago, IL 60612, USADepartment of Anatomy and Cell Biology, University of Illinois at Chicago, 808 South Wood Street, Rm 578 MC 512, Chicago, IL 60612, USADepartment of Anatomy and Cell Biology, University of Illinois at Chicago, 808 South Wood Street, Rm 578 MC 512, Chicago, IL 60612, USADepartment of Applied Physics and Materials Science, Northern Arizona University, Flagstaff, AZ 86011, USADepartment of Anatomy and Cell Biology, University of Illinois at Chicago, 808 South Wood Street, Rm 578 MC 512, Chicago, IL 60612, USADepartment of Anatomy and Cell Biology, University of Illinois at Chicago, 808 South Wood Street, Rm 578 MC 512, Chicago, IL 60612, USALight sheet microscopy in live cells requires minimal excitation intensity and resolves three-dimensional (3D) information rapidly. Lattice light sheet microscopy (LLSM) works similarly but uses a lattice configuration of Bessel beams to generate a flatter, diffraction-limited z-axis sheet suitable for investigating subcellular compartments, with better tissue penetration. We developed a LLSM method for investigating cellular properties of tissue in situ. Neural structures provide an important target. Neurons are complex 3D structures, and signaling between cells and subcellular structures requires high resolution imaging. We developed an LLSM configuration based on the Janelia Research Campus design or in situ recording that allows simultaneous electrophysiological recording. We give examples of using LLSM to assess synaptic function in situ. In presynapses, evoked Ca<sup>2+</sup> entry causes vesicle fusion and neurotransmitter release. We demonstrate the use of LLSM to measure stimulus-evoked localized presynaptic Ca<sup>2+</sup> entry and track synaptic vesicle recycling. We also demonstrate the resolution of postsynaptic Ca<sup>2+</sup> signaling in single synapses. A challenge in 3D imaging is the need to move the emission objective to maintain focus. We have developed an incoherent holographic lattice light-sheet (IHLLS) technique to replace the LLS tube lens with a dual diffractive lens to obtain 3D images of spatially incoherent light diffracted from an object as incoherent holograms. The 3D structure is reproduced within the scanned volume without moving the emission objective. This eliminates mechanical artifacts and improves temporal resolution. We focus on LLS and IHLLS applications and data obtained in neuroscience and emphasize increases in temporal and spatial resolution using these approaches.https://www.mdpi.com/2313-433X/9/6/121fluorescence microscopylattice light sheet holographyincoherent lattice light sheetelectrophysiology and simultaneous imagingin situ recordingsynaptic function |
| spellingShingle | Mariana Potcoava Donatella Contini Zachary Zurawski Spencer Huynh Christopher Mann Jonathan Art Simon Alford Live Cell Light Sheet Imaging with Low- and High-Spatial-Coherence Detection Approaches Reveals Spatiotemporal Aspects of Neuronal Signaling Journal of Imaging fluorescence microscopy lattice light sheet holography incoherent lattice light sheet electrophysiology and simultaneous imaging in situ recording synaptic function |
| title | Live Cell Light Sheet Imaging with Low- and High-Spatial-Coherence Detection Approaches Reveals Spatiotemporal Aspects of Neuronal Signaling |
| title_full | Live Cell Light Sheet Imaging with Low- and High-Spatial-Coherence Detection Approaches Reveals Spatiotemporal Aspects of Neuronal Signaling |
| title_fullStr | Live Cell Light Sheet Imaging with Low- and High-Spatial-Coherence Detection Approaches Reveals Spatiotemporal Aspects of Neuronal Signaling |
| title_full_unstemmed | Live Cell Light Sheet Imaging with Low- and High-Spatial-Coherence Detection Approaches Reveals Spatiotemporal Aspects of Neuronal Signaling |
| title_short | Live Cell Light Sheet Imaging with Low- and High-Spatial-Coherence Detection Approaches Reveals Spatiotemporal Aspects of Neuronal Signaling |
| title_sort | live cell light sheet imaging with low and high spatial coherence detection approaches reveals spatiotemporal aspects of neuronal signaling |
| topic | fluorescence microscopy lattice light sheet holography incoherent lattice light sheet electrophysiology and simultaneous imaging in situ recording synaptic function |
| url | https://www.mdpi.com/2313-433X/9/6/121 |
| work_keys_str_mv | AT marianapotcoava livecelllightsheetimagingwithlowandhighspatialcoherencedetectionapproachesrevealsspatiotemporalaspectsofneuronalsignaling AT donatellacontini livecelllightsheetimagingwithlowandhighspatialcoherencedetectionapproachesrevealsspatiotemporalaspectsofneuronalsignaling AT zacharyzurawski livecelllightsheetimagingwithlowandhighspatialcoherencedetectionapproachesrevealsspatiotemporalaspectsofneuronalsignaling AT spencerhuynh livecelllightsheetimagingwithlowandhighspatialcoherencedetectionapproachesrevealsspatiotemporalaspectsofneuronalsignaling AT christophermann livecelllightsheetimagingwithlowandhighspatialcoherencedetectionapproachesrevealsspatiotemporalaspectsofneuronalsignaling AT jonathanart livecelllightsheetimagingwithlowandhighspatialcoherencedetectionapproachesrevealsspatiotemporalaspectsofneuronalsignaling AT simonalford livecelllightsheetimagingwithlowandhighspatialcoherencedetectionapproachesrevealsspatiotemporalaspectsofneuronalsignaling |