A5020940486- Photoreceptor and optogenetics research
- Advanced Fluorescence Microscopy Techniques
- bioluminescence and chemiluminescence research
- Retinal Development and Disorders
- Advanced biosensing and bioanalysis techniques
- Cell Image Analysis Techniques
- Ion Transport and Channel Regulation
- Ion channel regulation and function
- Zebrafish Biomedical Research Applications
- Virus-based gene therapy research
- Neuroscience and Neural Engineering
- CCD and CMOS Imaging Sensors
- RNA and protein synthesis mechanisms
- Neuroscience and Neuropharmacology Research
- Hepatitis B Virus Studies
- Birth, Development, and Health
- Nanoplatforms for cancer theranostics
- Effects of Environmental Stressors on Livestock
- Cervical Cancer and HPV Research
- Click Chemistry and Applications
- Pharmacological Effects and Assays
- Cancer Research and Treatments
- Photoacoustic and Ultrasonic Imaging
- Single-cell and spatial transcriptomics
- Optical Imaging and Spectroscopy Techniques
Massachusetts Institute of Technology
2022-2024
McGovern Institute for Brain Research
2020-2024
Institute of Quality Standards and Testing Technology for Agro Products
2024
Chinese Academy of Agricultural Sciences
2024
The University of Tokyo
2024
University of Alberta
2017-2022
Howard Hughes Medical Institute
2020
Abstract The glymphatic movement of fluid through the brain removes metabolic waste 1–4 . Noninvasive 40 Hz stimulation promotes neural activity in multiple regions and attenuates pathology mouse models Alzheimer’s disease 5–8 Here we show that multisensory gamma influx cerebrospinal efflux interstitial cortex 5XFAD model disease. Influx was associated with increased aquaporin-4 polarization along astrocytic endfeet dilated meningeal lymphatic vessels. Inhibiting clearance abolished removal...
Abstract Potassium ion (K + ) homeostasis and dynamics play critical roles in biological activities. Here we describe three genetically encoded K indicators. KIRIN1 (potassium ( i on r atiometric dicator) KIRIN1-GR are Förster resonance energy transfer (FRET)-based indicators with a bacterial binding protein (Kbp) inserting between the fluorescent FRET pairs mCerulean3/cp173Venus Clover/mRuby2, respectively. GINKO1 g reen dicator of for o ptical imaging) is single protein-based indicator...
The upgraded version of intelligent image-activated cell sorting (iIACS) has enabled higher-throughput and more sensitive image-based single live cells from heterogeneous populations.
Genetically encoded calcium ion (Ca2+) indicators (GECIs) are indispensable tools for measuring Ca2+ dynamics and neuronal activities in vitro vivo. Red fluorescent protein (RFP)-based GECIs have inherent advantages relative to green protein-based due the longer wavelength light used excitation. Longer is associated with decreased phototoxicity deeper penetration through tissue. GECI can also enable multicolor visualization blue- or cyan-excitable fluorophores. Here we report development,...
Near-infrared (NIR) genetically encoded calcium ion (Ca2+) indicators (GECIs) can provide advantages over visible wavelength fluorescent GECIs in terms of reduced phototoxicity, minimal spectral cross talk with light excitable optogenetic tools and probes, decreased scattering absorption mammalian tissues. Our previously reported NIR GECI, NIR-GECO1, has these but also several disadvantages including lower brightness limited fluorescence response compared to state-of-the-art GECIs, when used...
Molecular signals interact in networks to mediate biological processes. To analyze these networks, it would be useful image many at once, the same living cell, using standard microscopes and genetically encoded fluorescent reporters. Here, we report temporally multiplexed imaging (TMI), which uses proteins with different clocklike properties—such as reversibly photoswitchable switching kinetics—to represent cellular signals. We linearly decompose a brief (few-second-long) trace of...
Glutamate is one of the 20 common amino acids and utmost importance for chemically mediated synaptic transmission in nervous systems. To expand color palette genetically encoded indicators glutamate, we used protein engineering to develop a red intensity-based glutamate-sensing fluorescent reporter (R-iGluSnFR1). Manipulating topology R-iGluSnFR1, previously reported green indicator, led development noncircularly permutated (ncp) variants. R- Rncp-iGluSnFR1 display glutamate affinities 11 μM...
For over 20 years, genetically encoded Ca2+ indicators have illuminated dynamic signaling activity in living cells and, more recently, whole organisms. We are just now beginning to understand how they work. Various fluorescence colors of these been developed, including red. Red ones promising because longer wavelengths light scatter less tissue, making it possible image deeper. They engineered from a red fluorescent protein that is circularly permuted and fused Ca2+-sensing domain. When...
Abstract Fluorescent genetically encoded calcium ion indicators (GECIs) enable Ca 2+ dynamics to be monitored in a diverse array of cell types and tissues. One drawback green fluorescent GECIs, such as the widely used GCaMP6, is that blue wavelengths light excite GECI also activate optogenetic actuators channelrhodopsins. Accordingly, it particularly challenging simultaneously use both GECIs control image signaling. Bioluminescence an alternative imaging modality circumvents this problem by...
Abstract Far‐red and near‐infrared (NIR) genetically encoded calcium ion (Ca 2+ ) indicators (GECIs) are powerful tools for in vivo multiplexed imaging of neural activity cell signaling. Inspired by a previous report to engineer far‐red fluorescent protein (FP) from biliverdin (BV)‐binding NIR FP, we have developed GECI, designated iBB‐GECO1, previously reported GECI. iBB‐GECO1 exhibits relatively high molecular brightness, an inverse response Ca with Δ F / min = −13, near‐optimal...
High-speed wide-field fluorescence microscopy has the potential to capture biological processes with exceptional spatiotemporal resolution. However, conventional cameras suffer from low signal-to-noise ratio at high frame rates, limiting their ability detect faint fluorescent events. Here, we introduce an image sensor where each pixel individually programmable sampling speed and phase, so that pixels can be arranged simultaneously sample a ratio. In high-speed voltage imaging experiments,...
Abstract Near-infrared (NIR) genetically-encoded calcium ion (Ca 2+ ) indicators (GECIs) can provide advantages over visible wavelength fluorescent GECIs in terms of reduced phototoxicity, minimal spectral cross-talk with visible-light excitable optogenetic tools and probes, decreased scattering absorption mammalian tissues. Our previously reported NIR GECI, NIR-GECO1, has these but also several disadvantages including lower brightness limited fluorescence response compared to...
Abstract Potassium ion (K + ) homeostasis and dynamics play critical roles in regulating various biological activities, the ability to monitor K spatial-temporal is understanding these functions. Here we report design characterization of a Förster resonance energy transfer (FRET)-based genetically encoded indicator, KIRIN1, constructed by inserting bacterial cytosolic binding protein (Kbp) between fluorescent (FP) FRET pair, mCerulean3 cp173Venus. Binding induces conformational change Kbp,...
Abstract Molecular signals interact to mediate diverse biological computations. Ideally one would be able image many at once, in the same living cell, reveal how they work together. Here we report temporally multiplexed imaging (TMI), which uses clocklike properties of fluorescent proteins enable different cellular represented by temporal fluorescence codes. Using photoswitchable represent signals, can linearly decompose a brief movie fluctuations given into sum fluctuation traces each...
Abstract Citrate esters have become the main alternatives to traditional plasticizers in food packaging materials. However, there is a lack of understanding their toxic effects, particularly combined effects and inner mechanisms has not been well studied. Our group pioneered study on toxicity tributyl citrate (TBC) acetyl triethyl (ATEC), two commonly co‐used The results showed that exposure TBC ATEC can decrease viability HepG2 cells dose dependent manner. When mixtures exposed cells, they...
Fourteen collaborating laboratories assayed maleic hydrazide (MH), 6-hydroxypyridazin-3(2H)-one, in technical and formulated products by reversed-phase liquid chromatography (LC) with sulfanilic acid as an internal standard. The active MH the samples (6 lots) ranged from 16% (expressed potassium salt) to 98% (MH technical). A small amount of 1 M KOH was added analytical standards create salt analyte which is soluble water. Test were extracted water containing standard before analysis LC on a...
High-speed wide-field fluorescence microscopy has the potential to capture biological processes with exceptional spatiotemporal resolution. However, conventional cameras suffer from low signal-to-noise ratio at high frame rates, limiting their ability detect faint fluorescent events. Here, we introduce an image sensor where each pixel individually programmable sampling speed and phase, so that pixels can be arranged simultaneously sample a ratio. In high-speed voltage imaging experiments,...
Abstract Genetically encoded Ca 2+ indicators (GECIs) are widely used to illuminate dynamic signaling activity in living cells and tissues. Various fluorescence colors of GECIs available, including red. Red promising because longer wavelengths light scatter less tissue, making it possible image deeper. They engineered from a circularly permuted red fluorescent protein fused sensing domain, calmodulin calmodulin-binding peptide. A conformational change the domain upon binding causes intensity...
Abstract Genetically-encoded calcium ion (Ca 2+ ) indicators (GECIs) are indispensable tools for measuring Ca dynamics and neuronal activities in vitro vivo. Red fluorescent protein (RFP)-based GECIs enable multicolor visualization with blue or cyan-excitable fluorophores combined use optogenetic actuators. Here we report the development, structure, validation of a new red indicator, K-GECO1, based on circularly permutated RFP derived from sea anemone Entacmaea quadricolor. We characterized...