Fluorescent Sensors for Imaging Interstitial Calcium

Valiente-Gabioud, Ariel A.; Suner, Ines Garteizgogeascoa; Idziak, Agata; Fabritius, Arne; Angibaud, Julie; Basquin, J.; Nägerl, U. Valentin; Singh, Sumeet; Griesbeck, Oliver

doi:10.1101/2023.03.23.533956

Cited by 2 publications

(2 citation statements)

References 51 publications

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“…Super-resolution shadow imaging has further been used consecutively with conventional confocal imaging of intracellular calcium transients as reported by the GCaMP6f genetically encoded calcium indicator ( Velicky et al, 2023 ; Figure 5F ) and a newly introduced low-affinity genetically encoded calcium indicator, GreenT, that localize to the cell membrane and reportedly respond to changes in interstitial calcium ( Valiente-Gabioud et al, 2023 ).…”

Section: Combining Shadow Imaging With Functional Optical Approachesmentioning

confidence: 99%

Fluorescence microscopy shadow imaging for neuroscience

Inavalli,

Puente Muñoz,

Draffin

et al. 2024

Front. Cell. Neurosci.

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Fluorescence microscopy remains one of the single most widely applied experimental approaches in neuroscience and beyond and is continuously evolving to make it easier and more versatile. The success of the approach is based on synergistic developments in imaging technologies and fluorophore labeling strategies that have allowed it to greatly diversify and be used across preparations for addressing structure as well as function. Yet, while targeted labeling strategies are a key strength of fluorescence microscopy, they reciprocally impose general limitations on the possible types of experiments and analyses. One recent development that overcomes some of these limitations is fluorescence microscopy shadow imaging, where membrane-bound cellular structures remain unlabeled while the surrounding extracellular space is made to fluoresce to provide a negative contrast shadow image. When based on super-resolution STED microscopy, the technique in effect provides a positive image of the extracellular space geometry and entire neuropil in the field of view. Other noteworthy advantages include the near elimination of the adverse effects of photobleaching and toxicity in live imaging, exhaustive and homogeneous labeling across the preparation, and the ability to apply and adjust the label intensity on the fly. Shadow imaging is gaining popularity and has been applied on its own or combined with conventional positive labeling to visualize cells and synaptic proteins in their parenchymal context. Here, we highlight the inherent limitations of fluorescence microscopy and conventional labeling and contrast these against the pros and cons of recent shadow imaging approaches. Our aim is to describe the brief history and current trajectory of the shadow imaging technique in the neuroscience field, and to draw attention to its ease of application and versatility.

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Section: Combining Shadow Imaging With Functional Optical Approachesmentioning

confidence: 99%

Fluorescence microscopy shadow imaging for neuroscience

Inavalli,

Puente Muñoz,

Draffin

et al. 2024

Front. Cell. Neurosci.

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“…Nevertheless, millimolar affinity sensors can prove useful for monitoring extracellular calcium dynamics, 45 or visualizing calcium flux inside subcellular compartments such as the endoplasmic reticulum. [46][47][48] Overall, 13-HaloCaMP1a provided the largest photoacoustic signal in the Ca 2+ -bound state (Figure 4d), and 18-HaloCaMP1b showed the highest sensitivity, with ∆PA/PA0 = 7.5 (Figure 4e).…”

Section: Design and Properties Of Photoacoustic Calcium Sensorsmentioning

confidence: 99%

Chemigenetic far-red labels and Ca²⁺indicators optimized for photoacoustic imaging

Cook,

Kaydanov,

Ugarte-Uribe

et al. 2024

Preprint

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Photoacoustic imaging is an emerging modality with significant promise for biomedical applications such as neuroimaging, owing to its capability to capture large fields of view, deep inside complex scattering tissue. However, the widespread adoption of this technique has been hindered by a lack of suitable molecular reporters for this modality. In this work, we introduce chemigenetic labels and calcium sensors specifically tailored for photoacoustic imaging, using a combination of synthetic dyes and HaloTag-based self-labelling proteins. We rationally design and engineer far-red "acoustogenic" dyes, showing high photoacoustic turn-ons upon binding to HaloTag, and develop a suite of tunable calcium indicators based on these scaffolds. These first-generation photoacoustic reporters show excellent performance in tissue-mimicking phantoms, with the best variants outperforming existing sensors in terms of signal intensity, sensitivity and photostability. We demonstrate the application of these ligands for labelling HaloTag-expressing neurons in mouse brain tissue, producing strong, specifically targeted photoacoustic signal, and provide a first example of in vivo labelling with these chemigenetic photoacoustic probes. Together, this work establishes a new approach for the design of photoacoustic reporters, paving the way towards deep tissue functional imaging.

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Fluorescent Sensors for Imaging Interstitial Calcium

Cited by 2 publications

References 51 publications

Fluorescence microscopy shadow imaging for neuroscience

Fluorescence microscopy shadow imaging for neuroscience

Chemigenetic far-red labels and Ca²⁺indicators optimized for photoacoustic imaging

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Fluorescent Sensors for Imaging Interstitial Calcium

Cited by 2 publications

References 51 publications

Fluorescence microscopy shadow imaging for neuroscience

Fluorescence microscopy shadow imaging for neuroscience

Chemigenetic far-red labels and Ca2+indicators optimized for photoacoustic imaging

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Chemigenetic far-red labels and Ca²⁺indicators optimized for photoacoustic imaging