...cereblon Optimized mScarlet-based tags for live imaging in C. elegans CRISPR/Cas9 for gene disruption in... Anti-V5 [IPI-SV5-Pk1] in a western blot assay. Image reused from Anderson et al., 2024 under CC BY license...ubiquitination substrate Ikaros (IKZF1ZF2) (B). Image adapted from Kroupova et al., 2024 under CC BY license... 39406745. Optimizing mScarlet tags for live imaging in C. elegans By Alyssa Neuhaus Red fluorescent...stages of embryonic development. Representative images of single focal planes through the middle of each...mScarlet-I3 constructs. Scale bar for all panels: 20 µm. Image reused from Cao et al., 2024 under CC-BY license...scenarios requiring rapid maturation and prolonged imaging with minimal background.   Find these C. elegans...

...of microscopy, where image processing algorithms for analyzing time-lapse images, quantitatively evaluating...CARE), segmentation of imaging data (ILASTIK), or the popular plugin-based image process framework FIJI...almost all disciplines. In biology, innovative new imaging approaches are constantly being developed which...through a microscope. Lower left panel shows an image of differentiating monocyte with planes and hardware...cutting-edge research to everybody by relying on tailored image processing and low-cost optical setups. References...Manu Prakash, "Squid: Simplifying Quantitative Imaging Platform Development and Deployment." bioRxiv. ...-cost 3D-printed open standard for microscopic imaging. Nat Commun 11, 5979 (2020). https://doi.org/10.1038...

...freeware.    Fig. 4: Images of 12 visual barcodes in use in A375 cell line. Image from Kaufman, et al,...Schematic showing the workflow for the BLITZ system. Image from Xiong et al. 2021.    BLITZ uses the biotin... of a CAG-driven mNeonGreen construct in mouse. Image from Goertsen D, et al., 2022.  Find M-CREATE... dCas9-based gene editor using microbial SSAPs. Image from Wang, et al, 2022.  Find dCas9-SSAP plasmids...clonal multiplexing by: Rachel Leeson Multiplexed images in live cells remains a challenge due to multiple...barcodes for clonal multiplexing of cells in live imaging application. Each visual barcode consists of a ... they were able to develop a 12-plex multiplex imaging system. Mixing the subclones within a single well...

...neurons, imaged 14 days post-transfection, excited at 505 nm (right). Scalebar represents 100 µm. Image from...and pain. Once these are completed, Hubbard will image these neurons using GCaMP7, measuring calcium signaling...disorder,” says Hubbard. Figure 3: 20x confocal image of the periaqueductal gray (PAG), with red neurons...expressing tdTomato and green neurons expressing cFos. Image from Elizabeth Hubbard.   Haoyi Qiu: Pain response...of the spinal cord. Created with BioRender.com. Image from Haoyi Qiu.   He Tian: Directed evolution...high-performance sensors to enable multiplexed imaging of neurotransmitter, calcium, and membrane voltage...Citrine-tagged voltage indicator expressed in mouse cortex. Image from He Tian. Congrats again to all the recipients...

...NST. (A-C) Low-magnification images showing viral spread. (D-F)  High-magnification images immuno-stained...methods and image descriptions, a minimum n = 2, and well labeled high- and low-magnification images demonstrating...immuno-stained against dopamine beta-hydroxylase (DBH, magenta) to label norepinephrine (NE) expressing neurons and...

...article in the BBC Wildlife Magazine. It is easy to see how these incredible images are attracting so much ...interest for its stunning images that is has been featured in Time magazine . Other smaller competitions...an aesthetic level as well. Beautiful microscope images which traditionally languished in the pages of ...within flowers with the use of x-rays. His haunting images have been enchanting people for years and have ...featured in the National Geographic and Smithsonian Magazine, where the editor stated “Albert G. Richards, ...

... challenges in 2-photon in-vivo imaging of voltage fluctuations. Image credit: Liu Z, et al. Cell. 2022...found an indicator that is optimized for 2-photon imaging of voltages. This includes monitoring of fast bursts...in mouse. C) Action potential frequencies with (magenta) and without (black) photoactivation of HcKCR1 ...top) and with (bottom) photoactivation of HcKCR1. Images adapted from Govorunova et al. 2022.     HcKCRs...know. We’d love to hear from you!   Fig. 3: Image of Addgene’s western blot for the myc-tagged protein...on the Anti-c-Myc [9E10] page for more details. Image credit: Addgene.   Find anti-c-Myc antibody ...

...demonstrated through living images created by the team (Fig. 2). These images are based on the concept of...bacteriograph was created using 160 individual images stitched together. Image from Romano et al., 2021 with permission...bacteriographs created about a decade ago. To create these images, the scientists used a strain transformed with ...mask over the plate containing the pattern of the image, and shined blue light over the plate. Areas on ...areas that are exposed will fluoresce. With these images, you can see just how spatially precise the activation...

... our blog. Image: The final and required step requires submission of an experimental image result that... with a caption. For microscopy images, please include channel imaged, a color legend for probes/dyes/...promoter, or tool type and each entry contains an image along with key experimental details (e.g. titer,...and objective used. If you did not generate the image yourself and it is under copyright, you will need...owner and will have the opportunity to cite the image to give proper attribution.  We hope that this ...

...from traditional imaging to FRET and stochastic single-molecule super resolution imaging. mNeonGreen also...yellow fluorescent protein wavelengths, it can be imaged either with standard green fluorescent protein ...2.6-fold increase in signal relative to mCerulean imaged with a standard CFP set. mTFP1 can be used as a...donor to mCitrine and mOrange or for live cell imaging of fusions and FRET reporters. mTFP1 is also insensitive...characterization and applications in fluorescence imaging." Biochemical Journal 400.3 (2006): 531-540. PubMed...determinants of color and applications in fluorescence imaging." BMC biology 6.1 (2008): 13. PubMed PMID: 18325109...

...and green channels overlaid on a bright-field image. Image from Ma et al, 2016. Using dCas9 to visualize...represents a gRNA targeting a specific chromosome. This image represents the composite of fluorescent measurements...Red boxB-boxB Cyan MS2-PP7 Yellow PP7-boxB Magenta boxB-MS2 White boxB-MS2-PP7 Despite the success...will produce a secondary color cyan, yellow, or magenta, depending on the hairpin combination bringing ...different hairpin sequences produce yellow, cyan, or magenta light. A gRNA with all 3 distinct hairpin sequences...  References 1. Chen, Baohui, et al. “Dynamic Imaging of Genomic Loci in Living Human Cells by an Optimized...

...integration, producing a second copy of the sgRNAs. Image reused from Walton et al. 2024 under a CC-BY-NC-...editing efficiencies at different loci in U2OS cells. Image adapted from Yan et al. 2024 under a CC-BY license...against vinculin as a loading control (right panel). Image from Addgene. These antibodies are recombinant...conjugated to a JaneliaFluor dye emitting at 525 nm. Image reused from https://www.janelia.org/open-science...Cre-dependent AAV1 vectors and are suitable for extended imaging of neurons in vivo. Because they are negative-going...    Figure 5: A) Representative confocal images showing GFP expression from ssAAV5s with hIBA1 ...splice acceptor site; pA, polyadenylation signal. Image reused from Riedmayr et al. 2023 under a CC-BY license...

...2015) or the UnaG protein that binds bilirubin (Kumagai, 2013). Next to these naturally occurring co-factor..., et al. "LOV-based reporters for fluorescence imaging." Current opinion in chemical biology 27 (2015)...2015): 39-45. PubMed PMID: 26087123. 4. Kumagai, Akiko, et al. "A bilirubin-inducible fluorescent protein...11. Várnai, Péter, and Tamas Balla. "Live cell imaging of phosphoinositides with expressed inositide binding...PMID: 18155467. 13. Chen, Baohui, et al. "Dynamic imaging of genomic loci in living human cells by an optimized...Nano-lanterns for real-time multicolor luminescence imaging." Proceedings of the National Academy of Sciences..., Satoshi, et al. "Single-cell bioluminescence imaging of deep tissue in freely moving animals." Science...

...specific subcellular regions, laser ablation can damage other cellular structures, and genetic mutations...body. B) Schematic of Opto-katanin’s use in cells. Images adapted from Meiring et al., 2022, under CC-BY ... Opto-RhoGEF design and activity. B) OptoTIAM (magenta) expressed in a monolayer of endothelial cells,...after blue light illumination. Scale bar 25 µm. Images adapted from Mahlandt et al., 2023, under CC-BY...protein vesicle trafficking via CRY2-CIB1 clustering. Image adapted from Aljiboury et al., 2023, under CC-BY...expressing plasma membrane-targeted SuperPLDx30 (magenta) and PA probe GFP-PASS (green). After 30 minutes...colocalizing with SuperPLDx30. Scale bar 10 µm. Images adapted from Tei et al. 2022, bioRxiv, under a ...

...and consisting of the fusion proteins N713-pMag and nMagHigh1-C714, this new system had both low background... system by modifying paCas9-1 using nMagC714 instead of nMagHigh1-C714, generating paCas9-2. This change...figure below. Their most successful design utilized Magnet photoswitchable proteins derived from the fungal...Additionally, high intensity exposure can cause DNA damage. Far-red light, however, can penetrate 5 mm past... be used to identify specific genomic loci for imaging techniques. Since expression of dCas9 can potentially...gene transcription Nihongaki et al., 2015 paCas9 Magnet proteins from N. crassa Split Cas9, Split Cas9 ...

...bacteria CRMAGE system combines CRISPR and recombineering-based MAGE The Nielsen lab’s CRMAGE system is...Pedersen LE, Sommer MOA, Nielsen AT (2016) CRMAGE: CRISPR Optimized MAGE Recombineering. Sci Rep 6: . https:/...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...

...lens which magnifies the image, then the ocular lens, where the image is further magnified before it finally...10x magnification. To determine the final magnification of your image, multiply the magnification of your...now) is the magnification power, such as 4x, 10x, or 20x. A 4x objective magnifies the image four times...properties of a microscope are magnification (the ability to make an image larger) and resolution (the ability...your eyes. These lenses determine the magnification of the image and the resolution your microscope can...10x objective magnifies it 10 times, and so on. The ocular lens also provides magnification and the power...also 10x, you would actually be seeing a 100x magnified image of that object! Equipment Light Microscope ...

...depends on both the biological system as well as the imaging method, we recommend basing your selection on a...application. The three key properties for live cell imaging that will be treated in this series of posts are...experimental conditions that are related to the imaging strategy and the sample. Practical brightness A...mammalian cells is to tag an endogenous gene. By imaging cells (or tissues) that produce an endogenous protein... be used to verify performance under different imaging conditions (e.g. different emission filters) or...Choosing Your Fluorescent Proteins for Multicolor Imaging Learn about Fluorescent Protein Biosensors Additional...

...this post: Anti-FLAG Frankenbodies for modular imaging of cellular targets Optimized genetic screening...and editing Anti-FLAG Frankenbodies for modular imaging of cellular targets by: Mike Lacy Adding to their...a variety of applications, including live-cell imaging, single-molecule tracking, immunolabeling of fixed... be streamlined into a single array for Cas12a. Image from DeWeirdt, et al. 2021 To validate this ...expressed and coexpressed proteins run as ladders. Image from Santilli, et al. 2021 Plasmids are used...of CasMINI for genome engineering applications. Image from Xiaoshu, et. al. 2021.   Created by the...

...antibody specificity via a colorimetric reaction. Image from Boguszewska et al., 2019.   Competitive...antibody specificity via a colorimetric reaction. Image from Boguszewska et al., 2019.       Direct ...antibody specificity via a colorimetric reaction. Image from Boguszewska et al., 2019.   Indirect ELISA...antibody specificity via a colorimetric reaction. Image from Boguszewska et al., 2019.   Detecting the...this line to calculate the protein concentration. Image from Jagarlamudi et al., 2015.   The maximum...Urbaniak, S. et al. Review: immunoassays in DNA damage and instability detection. Cell. Mol. Life Sci....