Single Photon Kilohertz Frame Rate Imaging of Neural Activity

Tian, Tian; Yuan, Yifang; Mitra, Srinjoy; Gyöngy, István; Nolan, Matthew F.

doi:10.1002/advs.202203018

Cited by 4 publications

(3 citation statements)

References 30 publications

(53 reference statements)

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“…46,48,49 The use of modern GEVIs in combination with modern microscopic technologies makes it possible to combine subcellular spatial resolution with millisecond resolution from identified neuronal populations in living brain slices. 50,51 For transient expression, indicators can be injected directly into the animal's brain using lentiviral or adenovirus vectors. An alternative way is to obtain transgenic animals expressing these indicators under the control of tissue-specific or constitutive promoters.…”

Section: Genetically Encoded Indicators Of Neural Activitymentioning

confidence: 99%

Neurophotonic methods in approach to in vivo animal epileptic models: Advantages and limitations

Tsytsarev,

Sopova,

Leonova

et al. 2024

Epilepsia

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Neurophotonic technology is a rapidly growing group of techniques that are based on the interactions of light with natural or genetically modified cells of the neural system. New optical technologies make it possible to considerably extend the tools of neurophysiological research, from the visualization of functional activity changes to control of brain tissue excitability. This opens new perspectives for studying the mechanisms underlying the development of human neurological diseases. Epilepsy is one of the most common brain disorders; it is characterized by recurrent seizures and affects >1% of the world's population. However, how seizures occur, spread, and terminate in a healthy brain is still unclear. Therefore, it is extremely important to develop appropriate models to accurately explore the causal relationship of epileptic activity. The use of neurophotonic technologies in epilepsy research falls into two broad categories: the visualization of neural epileptic activity, and the direct optical influence on neurons to induce or suppress epileptic activity. An optogenetic variant of the classical kindling model of epileptic seizures, in which activatable cells are genetically defined, is called optokindling. Research is also underway concerning the application of neurophotonic techniques for suppressing epileptic activity, aiming to bring these methods into clinical practice. This review aims to systematize and describe new approaches that use combinations of different neurophotonic methods to work with in vivo models of epilepsy. These approaches overcome many of the shortcomings associated with classical animal models of epilepsy and thus increase the effectiveness of developing new diagnostic methods and antiepileptic therapy.

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Section: Genetically Encoded Indicators Of Neural Activitymentioning

confidence: 99%

Neurophotonic methods in approach to in vivo animal epileptic models: Advantages and limitations

Tsytsarev,

Sopova,

Leonova

et al. 2024

Epilepsia

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show abstract

“…The integration of ultra-high-speed cameras 7,8,21 and event cameras 22 holds promise for providing higher bandwidths to LFM. However, their current limitations in sensitivity and noise performance present challenges, especially for photon-starved applications like imaging genetically encoded voltage indicators (GEVIs).…”

Section: Introductionmentioning

confidence: 99%

Kilohertz volumetric imaging of in-vivo dynamics using squeezed light field microscopy

Wang,

Zhao,

Wagenaar

et al. 2024

Preprint

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Volumetric functional imaging of transient cellular signaling and motion dynamics poses a significant challenge to current microscopy techniques, primarily due to limitations in hardware bandwidth and the restricted photon budget within short exposure times. In response to this challenge, we present squeezed light field microscopy (SLIM), a computational imaging method that enables rapid detection of high-resolution three-dimensional (3D) light signals using only a single, low-format camera sensor area. SLIM pushes the boundaries of 3D optical microscopy, achieving over one thousand volumes per second across a large field of view of 550 μm in diameter and 300 μm in depth. Using SLIM, we demonstrated blood cell velocimetry across the embryonic zebrafish brain and in a free-moving tail exhibiting high-frequency swinging motion. The millisecond temporal resolution also enables accurate voltage imaging of neural membrane potentials in the leech ganglion. These results collectively establish SLIM as a versatile and robust imaging tool for high-speed microscopy applications.

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“…Some fruitful methods have been studied to suppress fluorescent noise, for example, using total internal reflection illumination, spinning disk confocal illumination, and light sheet microscopy. , The detector noises accompany each acquisition process, which are much less discussed. They are mainly determined by the performance of the detectors [charge-coupled devices (CCD) or complementary metal-oxide semiconductor (CMOS)]. , For SMLM with nanometer-level localization precision, especially under weak-signal conditions, detector noises can cause localization misalignments and signal losses, so their removal is indispensable. − However, due to the complexity of the detector noises, the multi-frame accumulate and average (ANA) , commonly used in experiments cannot efficiently eliminate these noises; therefore, the fluorescent signal extraction becomes less effective.…”

Section: Introductionmentioning

confidence: 99%

Enhancing Weak-Signal Extraction for Single-Molecule Localization Microscopy

Cheng

et al. 2022

J. Phys. Chem. A

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Single-molecule localization microscopy (SMLM) has been widely used in biological imaging due to its ultrahigh spatial resolution. However, due to the strategy of reducing photodamage to living cells, the fluorescence signals of emitters are usually weak and the detector noises become non-negligible, which leads to localization misalignments and signal losses, thus deteriorating the imaging capability of SMLM. Here, we propose an active modulation method to control the fluorescence of the probe emitters. It actually marks the emitters with artificial blinking character, which directly distinguishes weak signals from multiple detector noises. We demonstrated from simulations and experiments that this method improves the signal-to-noise ratio by about 10 dB over the non-modulated method and boosts the sensitivity of single-molecule localization down to −4 dB, which significantly reduces localization misalignments and signal losses in SMLM. This signal-noise decoupling strategy is generally applicable to the super-resolution system with versatile labeled probes to improve their imaging capability. We also showed its application to the densely labeled sample, showing its flexibility in super-resolution nanoscopy.

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Single Photon Kilohertz Frame Rate Imaging of Neural Activity

Cited by 4 publications

References 30 publications

Neurophotonic methods in approach to in vivo animal epileptic models: Advantages and limitations

Neurophotonic methods in approach to in vivo animal epileptic models: Advantages and limitations

Kilohertz volumetric imaging of in-vivo dynamics using squeezed light field microscopy

Enhancing Weak-Signal Extraction for Single-Molecule Localization Microscopy

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