...of the antibody binding “distinct imaged pattern.” The distinct imaged pattern should align with what the...contains secondary antibodies, allowing us to only image autofluorescence i.e., background noise. We then...and not from the background noise. If the sample images similarly to the negative control, that informs...on the antibody signal’s location. If a distinct imaged pattern does NOT align with the atlases/literature... own analysis of the background noise, distinct imaged pattern, and exposure times. To test the antibody...

...AC, BC, and ABC). Nielsen Lab E. coli CRMAGE Plasmids: The CRMAGE system is a fast, multiplexable method...12. Ronda, Carlotta, et al. (2016). "CRMAGE: CRISPR Optimized MAGE Recombineering." Sci Rep. 6:19452. PubMed...method that combines CRISPR and recombineering-based MAGE (Multiplex Automated Genome Engineering) technology...self-destruction" after induction with L-rhamnose and aTet. CRMAGE is much more efficient than traditional recombineering...simultaneously is possible with efficiency >70%. CRMAGE is an incredibly fast protocol, with only 5 hours...

...sea anemone's red color can be used for mimicry. Image: Bernard Spragg. Advantages of using chromoproteins...Chromoprotein expression from bacteria on agar plate. Image: Liljeruhm et al., 2018. For each chromoprotein... plasmids expressing amilCP and tsPurple genes. Image: Lazarus et al., 2019. One such lab is the Waldor...chromoprotein is expressed and gives the cell a magenta (amilCP) or purple (tsPurple) color. Though the... used as a FRET acceptor to mRuby2 or mScarlet. Image: Murakoshi et al., 2019. ShadowR is an orange...developed as an acceptor for fluorescence lifetime imaging microscopy-based Förster resonance energy transfer...

...Choosing the best PE6 variant for your application. Image reused form Doman and Pandey et al., 2023 under ...fusion and targeting components are interchangeable. Image from Strebinger et al., 2023, reused under CC-BY... trafficked from the “source” to “destination”. Images adapted from Lee and Cheah et al. 2023 bioRxiv ...GFP-iPAK4 within an hour. Analyzing the ticker tape images could recover transcriptional histories, accurate...labeled with HaloTag-iPAK4. Scale bar 10 µm. C) Images of iPAK4 fibers in cultured rat hippocampal neurons...induces cFos::GFP-iPAK4 at 3 or 6 h. Scale bar 5 µm. Images adapted from Lin et al., 2023 bioRxiv, under CC-BY-NC...

...immunofluorescence and indirect immunofluorescence work. Image created with BioRender.com   Indirect IF is ...access their target and that you will be able to image them in the end. Again, the methods you use will...time for this step as well. Preservation and Imaging Similar to Sample Preparation, this step can ...procedure with key considerations about each step. Image created with BioRender.com   The first couple.... the spectral properties match your available imaging instruments and the spectral properties do not ... pressing research questions, but also stunning images (am I the only one with IF results as artwork at...

...studies that involve surgery or imaging.  Rattus norvegicus. Image from Staffan Vilcans. Rat physiology...Idalia & Bernardo, 2017).  Escherichia coli. Image from NIAID. E. coli is often a part of the “molecular...mexicanum) Ambystoma mexicanum, aka the axolotl. Image from AJC1. The Mexican axolotl is one of the ...biomedical research.  Xenopus, aka African frog. Image from Holger Krisp. The frog genome has high synteny...adaptations.  Arabidopsis thaliana flowers. Image from BlueRidgeKitties. If you are working with...

...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, ...

...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...

... 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...

...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...

...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...

...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...

... 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. ...