...vivo. Merged green and red channel fluorescence images of C2C12 cells stably expressing pTRE-Tight-MitoTimer...pulsed with 2µg/ml of Doxycycline for 2 hours, and imaged on the Keyence BZ-9000 fluorescence microscope ...organelles with older (yellow to orange) protein. Image courtesy of Roberta Gottlieb. Laker et al., ...exhibit superior performance over APEX for both EM imaging and proximity-dependent proteomic mapping with ...which can be fused to proteins of interest for EM imaging, or targeted to subcellular regions or protein ...40.  Next-Gen Brainbow toolkit for neuronal imaging Joshua Sanes and his team at the Center for Brain...Brainbow toolkit for high-resolution fluorescent imaging of individual neurons. The technology generates...

...PsMOrange and mEos add another layer of complexity to imaging, as they allow one to monitor compartment/protein...subsequently confirmed that purified GFP could be imaged in the green state after excitation at 488 nm, ...the red state after excitation at 561 nm. This imaging process worked well with the common GFP variants...spreads through the cytosol via cytoplasmic strands. Image from Sattarzadeh et al., Sci Rep. 2015 CC BY 4.0...variety of other fluorescent proteins for your imaging needs. If you need an FP refresher, see our FP ...

...fluorophores for imaging Lee S, et al. (2013) PLoS ONE 8(7): e67902. Many imaging studies rely on...which nearly all domains of life repair genomic damage, specifically double strand breaks. Researchers...recommend optimal FP combinations for use in yeast imaging, categorized by specific filter sets and experimental...tagging vectors for your single or multi-color imaging experiments. Interested in epitope tags? Others...

...level provided by SunTag is so high, cells may be imaged under lower illumination settings, lowering photobleaching...phototoxicity concerns and extending the potential imaging time for this technique. The NIH Common Fund has...based method is a great step forward in live cell imaging of genomic loci. Ma et al. anticipate that the ...plasmids from this publication at Addgene. Dynamic imaging of genomic loci in living human cells by an optimized...amplification in gene expression and fluorescence imaging. Tanenbaum ME, Gilbert LA, Qi LS, Weissman JS, ...

... family of calcium sensors and can be used for imaging of small neuronal processes like dendrites and ...neurotransmitter in vertebrates, but methods for directly imaging GABA have been limited. Like calcium sensors, different...to monitor neuronal electrical activity through imaging. Since it’s illuminated by red/orange light, Archon1...resting fluorescence. It is useful for activity imaging of large populations of densely labeled neurons...resting fluorescence.  It is useful for activity imaging of large populations of densely-labeled neurons...

...in the CRISPR-pooled screening using sgRNAiBAR. Image from Zhu et al., 2019.  Wensheng Wei's lab at...indicator, GCaMP6f, in the Drosophila optic lobe. Image from Isaacman-Beck et al., 2019. Get started... stained with PI (pink) to indicate cell death. Image from Diessl et al., 2020. Find the GFP reporter...with the fluorescent protein fluorescing green. Image from Shin et al., 2019. Looking for an alternative...not cause structural deformation or photo-induced damage. These fusions can also function in high salt conditions...

...fiction. Two dueling magicians in the 1800s in England who help bring back actual magic (which had been lost...features a young female protagonist, some clever magic, and a satisfying conclusion? In this series, there...even more sinister. 1000 pages long. Incredibly imaginative, funny, bittersweet, and shocking at times."     ...ve read books across nearly every genre you can imagine: gothic, mystery, sci-fi, fantasy, pop-sci, true...

...external signals such as developmental cues and DNA damage. These signals activate a cascade of caspases, ...after proteolytic cleavage. Secondly, fluorescence imaging of living organisms can be challenging due to issues...array of multicolor reporters that will enable imaging of multiple proteases at once (Zhang et al., 2019...of a GFP-based fluorogenic caspase reporter for imaging apoptosis in vivo." Cell chemical biology 23.7 ... reporter by flipping a beta strand of GFP for imaging apoptosis in animals." Journal of the American ...

...likely already have a good idea of the basics of immunoimaging. A scientist conjugates an antibody with a signaling...will also be. This is the direct approach to immunoimaging assays. But if the direct approach isn’t giving...one of the most challenging components of any immunoimaging assay. Secondary antibodies, on the other hand... With the exception of flow cytometry, most immunoimaging assays do not allow for absolute quantitative...antibodies will find themselves able to approach immunoimaging assays more confidently, collect a broader array...

...fluorescent protein, producing fluorescent signal. Image modified from Feng et al. 2017 under CC-BY license... rate of p53 tetramerization in response to DNA damage controls the p53 response. Gaglia G, Lahav G. Mol...eukaryotes) A Near-Infrared BiFC Reporter for In Vivo Imaging of Protein-Protein Interactions. Filonov GS, Verkhusha...proteins as tags and biosensors for multiscale imaging. Shcherbakova et al. Nat Commun. 2016 Vladislav... allows exogenous labeling for single-molecule imaging TagBiFC technique allows long-term single-molecule...

...potential synthetic chromatin mediators and probes. Image from Priode et al., 2021.   The potential tools... between the sender cell and the receiver cell. Image from Tang et al., 2020.   G-baToN can also be...degradation and quantification of this depletion using imaging-based measurements (Ashley et al., 2021).   ...Schematic showing the auxin-inducible degron system. Image from Ashley et al., 2021.   The AID system relies...INTRSECT project. Check out the new additions.  Image calcium transients in PV interneurons with AAV-S5E2...

...and mVenus (bottom). Cells were imaged every 30 seconds for 3 hours. Image from Lee et al., 2020. SPOTlight...can eliminate competing mobile genetic elements. Image by Basem Al-Shayeb in the Doudna Lab. Scientists...SPOTlight was used to image over 3 million cell types expressing variants of Yellow Fluorescent Protein...require high photostability, such as time-lapse imaging or with high light power detection of molecules...

... homologous recombination (HR). Adapted from an image provided by Manuel Gonçalves; made with BioRender... the various cell types ranged from 18% to 65% (Maggio et al., 2014). An alternative approach to NHEJ ...subjected to scarless homology-directed genome editing (Maggio et al., 2014). Gonçalves said they attribute this...doi.org/10.1161/circresaha.115.304351 Holkers M, Maggio I, Henriques SFD, Janssen JM, Cathomen T, Gonçalves...516:423–427 . https://doi.org/10.1038/nature13902 Maggio I, Holkers M, Liu J, Janssen JM, Chen X, Gonçalves...

... used for recombinant antibody production is Komagataella phaffii (a.k.a  Pichia pastoris) because it ...biorender.com   Another method of harvesting uses magnetic beads. In this method, beads are incubated with...suspension. Antibodies bind ligands on the beads and a magnet is used to separate the beads from the media (Brechmann... and the concentration may be calculated using imaging software. Any bands on the gel beside the heavy... Chotteau V. Antibody capture process based on magnetic beads from very high cell density suspension. ...

...expressed. Cells are then cultured and FACS sorted. Image from Mateo Sanchez and Alice Ting. Sanchez MI...sequence derived either from p63(1-29) and Lck. Image from Chertkova et al., 2020. In their BioRxiv... adsorption chromatography (IMAC) using Ni-NTA magnetic beads. Workflow of nanobody expression and...genomic DNA with the help of the piggyBa transposase. Image from Hazelbaker et al., 2020. The lab tested ...wavelengths above 1,000 nm and enable two-color imaging with red fluorescence protein-based calcium sensors...

...photobleaching method and the fluorescence lifetime imaging microscopy (FLIM) method. Acceptor photobleaching...destroy the donor molecule. Fluorescence lifetime imaging microscopy method FLIM has been developed more ...acceptor. It should be noted that FLIM is a slower imaging method, limiting its use in many FRET experiments...interpreting data. FRET is 20 years old but as you can imagine not old-fashioned at all. With the development ...Fluorescence resonance energy transfer (FRET) microscopy imaging of live cell protein localizations. J Cell Biol...

...the fused XFP can be used to amplify the signal. Imaging endogenous fluorescence in live animals only permits...fluorophores and that can be combined with new imaging techniques.  References Cai, Dawen, et al. "Improved...17972876. Pan, Y. Albert, et al. "Multicolor Brainbow imaging in zebrafish." Cold Spring Harbor Protocols 2011.1...Addgene blog Read an introduction to live cell imaging Learn more about the origin of the Brainbow system...collection Learn about fluorescent tools for in vivo imaging ...

...trend in biology towards single-cell resolution imaging. Sanes and Lichtman endeavored to create a map ... these Brainbow constructs into mice. When they imaged the mouse brains, they found many more than 3-4...suffered from a number of limitations. First, the imaging of Brainbow mouse tissue was challenging due to...described in Hugo J. Snippert, et al. (2010) and the MAGIC Marker strategy described in Karine Loulier, et ...given section of brain by at least an order of magnitude. It is clear that further refinements of the Brainbow...

...months at 4 °C.  Pro tip! If you get a speckled image, your protein-based blocker is either degraded or...to be compatible with stripping and probing.   Imaging At the final step of your western, you’ll have ...cost, but require no consumables, produce digital images, reach their limit of detection quickly (~1 minute...dynamic range that typically spans 2–5 orders of magnitude, which includes a wide dynamic range for fluorescent...Pro tip! You only need a resolution of ~150 µm to image a blot. This is because proteins diffuse semiradially...-based buffers Increased cost X-ray film imaging Increased time; increased consumables cost ...

... et al., 2008; Taylor et al., 2016; Hooper and Maguire, 2016). Recombinant adeno-associated viruses (rAAVs...1984; Van Duyne, 2001; Christenson Wick and Krook-Magnuson, 2018). Optogenetic FLEx vectors contain a strong... Channelrhodopsin-2 to multiple cell types for imaging and long-range circuit mapping." Journal of Neuroscience...Central PMCID: PMC4971574. Hooper, Andrew, and Jamie Maguire. "Characterization of a novel subtype of hippocampal...11340053. Wick, Zoé Christenson, and Esther Krook-Magnuson. "Specificity, versatility, and continual development...