Designing Large Field-of-view Two Photon Microscopy Using Optical Invariant Analysis

Bumstead, Jonathan R.; Park, Jasmine J.; Rosen, Isaac A.; Kraft, Andrew W.; Wright, Patrick W.; Reisman, Matthew D.; Côté, Daniel; Culver, Joseph P.

doi:10.1364/translational.2018.jth3a.60

Cited by 2 publications

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“…Furthermore, the “space-bandwidth product”, which signifies the FOV to spatial resolution ratio achieved under real measurement conditions (considering image sensor size and wavelength), reaches 4.3 × 10 8 (Materials and Methods, 34). Both of these metrics significantly outperform those reported in earlier studies on large FOV imaging methods (6–10,33). Although these metrics are essential indicators of our system’s performance, they do not solely determine its superiority over other large-FOV imaging systems.…”

Section: Discussionmentioning

confidence: 56%

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Volumetric trans-scale imaging of massive quantity of heterogeneous cell populations in centimeter-wide tissue and embryo

Ichimura,

Kakizuka,

Taniguchi

et al. 2023

Preprint

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We established a volumetric trans-scale imaging system with an ultra-large field-of-view (FOV) that enables simultaneous observation of millions of cellular dynamics in centimeter-wide three-dimensional (3D) tissues and embryos. Using a custom-made giant lens system with a magnification of 2× and a numerical aperture (NA) of 0.25, and a CMOS camera with more than 100 megapixels, we built a trans-scale scope AMATERAS-2, and realized fluorescence imaging with a lateral spatial resolution of approximately 1.2 µm across an FOV of approximately 1.5 × 1.0 cm2. The 3D resolving capability was realized through a combination of optical and computational sectioning techniques tailored for our low-power imaging system. We applied the imaging technique to 1.2 cm-wide section of mouse brain, and successfully observed various regions of the brain with sub-cellular resolution in a single FOV. We also performed time-lapse imaging of a 1-cm-wide vascular network during quail embryo development for over 24 hours, visualizing the movement of over 4.0 × 105vascular endothelial cells and quantitatively analyzing their dynamics. Our results demonstrate the potential of this technique in accelerating production of comprehensive reference maps of all cells in organisms and tissues, which contributes to understanding developmental processes, brain functions, and pathogenesis of disease, as well as high-throughput quality check of tissues used for transplantation medicine.

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Section: Discussionmentioning

confidence: 56%

“…The second index, the space-bandwidth product, considers the image sensor and wavelength while quantifying the ratio of actual resolution to FOV. These indices are calculated using the following simplified formulas, respectively (33). …”

Section: Methodsmentioning

confidence: 99%

Volumetric trans-scale imaging of massive quantity of heterogeneous cell populations in centimeter-wide tissue and embryo

Ichimura,

Kakizuka,

Taniguchi

et al. 2023

Preprint

View full text Add to dashboard Cite

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Designing a Compact, Low-Cost FRET Microscopy Platform for the Undergraduate Classroom

et al. 2020

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ABSTRACT Advances in fluorescent biosensors allow researchers to spatiotemporally monitor a diversity of biochemical reactions and secondary messengers. However, commercial microscopes for the specific application of Förster Resonance Energy Transfer (FRET) are prohibitively expensive to implement in the undergraduate classroom, owing primarily to the dynamic range required and need for ratiometric emission imaging. The purpose of this article is to provide a workflow to design a low-cost, FRET-enabled microscope and to equip the reader with sufficient knowledge to compare commercial light sources, optics, and cameras to modify the device for a specific application. We used this approach to construct a microscope that was assembled by undergraduate students with no prior microscopy experience that is suitable for most single-cell cyan and yellow fluorescent protein FRET applications. The utility of this design was demonstrated by measuring small metabolic oscillations by using a lactate FRET sensor expressed in primary mouse pancreatic islets, highlighting the biologically suitable signal-to-noise ratio and dynamic range of our compact microscope. The instructions in this article provide an effective teaching tool for undergraduate educators and students interested in implementing FRET in a cost-effective manner.

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Designing Large Field-of-view Two Photon Microscopy Using Optical Invariant Analysis

Cited by 2 publications

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Volumetric trans-scale imaging of massive quantity of heterogeneous cell populations in centimeter-wide tissue and embryo

Volumetric trans-scale imaging of massive quantity of heterogeneous cell populations in centimeter-wide tissue and embryo

Designing a Compact, Low-Cost FRET Microscopy Platform for the Undergraduate Classroom

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