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We narrowed to 839 results for: IGH@

Showing: 1 - 40 of 839 results

  1. New Norepinephrine Indicators: nLightG and nLightR

    Type
    Blog Post
    Published
    Dec. 12, 2023, 2:15 p.m.
    ... nLightR. A, Structural models of nLightG (left) and nLightR (right) generated using ColabFold (Mirdita...indicators: nLightG and nLightR (Kagiampaki et al., 2023).     Figure 1: Properties of nLightG and nLightR...al., 2023. nLight nLight sensors are based on alpha-1a adrenergic receptor; nLightG contains circularly...norepinephrine. nLight is 28-fold more selective than GRABNE, with both nLightG and nLightR showing significantly...experimental design. Find nLight plasmids here! Next steps While nLightG and nLightR greatly expand the toolbox... both nLightG and nLightR, and may soon have a red tool which matches the sensitivity of nLightG. Many...images of HEK293T cells and neurons expressing nLightG or nLightR before/after application of NE (10 μM) and...

  2. Light Sheet Fluorescence Microscopy

    Type
    Blog Post
    Published
    July 16, 2015, 2:30 p.m.
    ...based on using a thin plane (sheet) of light generated by sunlight to observe single gold particles with...V= ventral Developments in light sheet fluorescence microscopy Modern light sheet fluorescence microscopy...out-of-focus reflected light by using a pinhole.  However, the excitation light excites all of the fluorophores..., there is a drop in image brightness with increasing depth caused by light scattering and absorption....of this century has seen some major advances in light microscopy, particularly related to the neurosciences....  One of these advances in microscopy has been light sheet fluorescence microscopy (LSFM). The underlying...a fluorescent sample and captured the reflected light using a different objective lens oriented perpendicular...

  3. The Importance of High Titer for AAV Transductions

    Type
    Blog Post
    Published
    March 1, 2018, 2:11 p.m.
    ...experiments require higher gene expression and therefore higher titer than others. High titers are often ...vector to such a high titer when looking for effective gene expression. How do you achieve high titers in the...good ones into a cell to produce expression. The higher the titer, the better. AAV transport and transgene...transgene expression It’s clear that we need high titers, as contact between the cell and any individual AAV...the cell may not always be enough and therefore higher titer will be required. For example, retrograde...over an even larger area, thus necessitating even higher retrograde AAV titers for successful transduction...That means that once you drop below a certain (very high) threshold of viral particles per cell, you may ...

  4. Career Insights: Technical Support Specialist

    Type
    Blog Post
    Published
    Nov. 9, 2017, 2 p.m.
    ...the pressure to publish in high-impact journals, and the requirement for a high grant success rate. At this... this point, candidates are highly experienced but often have to start pursuing other options. While some...technical support specialist for 4 years. Read on for insights from my time in technical support including tips..., and resolve complaints about products. As you might expect, different companies produce different products... dialogue. This will give you a feeling of what might be involved in such a role. In addition, it can ...individual stories and get (sometimes refreshing) insights into how other scientists think and work. Most...to enhance the OD signals and then complain about high background. In many instances, variations of the...

  5. Lighting Up Cell Signaling with Photoswitchable Kinases

    Type
    Blog Post
    Published
    July 25, 2017, 1:20 p.m.
    ...fluorescence when exposed to 500 nm light.  Under 400 nm cyan light, Dronpa dimers dissociate, Dronpa goes...minute pulse of 500 nm light (turn on psRaf1) followed by 3 seconds of 400 nm light (turn off psRaf1). After...and use MEK1 as a model kinase. In this system, light is used to turn “on” and “off” MEK1 activity by ...is called psMEK1. When exposed to 500 nm violet light, the Dronpa dimers interact and lock MEK1 in an ... an inhibitor of choice, exposed to 500 nm cyan light to activate psMEK1, and the localization of ERK ...these same cells by 24-48 hours exposure to blue light also induced tail swelling in ~75% of worms (see...expressed in a tissue or organism that blue and violet light can penetrate. How can I try using photoswitchable...

  6. Antibodies 101: Selecting the Right Antibody

    Type
    Blog Post
    Published
    March 15, 2022, 1:15 p.m.
    ...data on the assay methods. You might find that while the website highlights a variation of your application...protein or protein region highly conserved across species? If the proteins are highly homologous between your...

  7. Antibodies 101: Choosing the Right Isotype

    Type
    Blog Post
    Published
    April 6, 2023, 3 p.m.
    ...antibodies have higher affinity and specificity than mouse monoclonal antibodies. Antibodies with higher affinities...monoclonal antibodies contain a single clone they have higher lot to lot specificity and tend towards more reproducible...When using a monoclonal antibody be sure to use a highly specific secondary antibody, rather than a broadly...at very low levels or your application requires a high degree of affinity, then species choice could very...may be the better choice for assays that require high affinity binding, such as immunoprecipitations. ...stain a target protein in mouse tissues, causes a high degree of background staining. This occurs because...antibody isotypes matter and helps you choose the right antibody for your application!  References and ...

  8. Is this the right place for me? 8 tactics for choosing a lab

    Type
    Blog Post
    Published
    Oct. 2, 2018, 12:56 p.m.
    ...for many years (at a very low salary I might add). You have a right and responsibility to choose a lab where...Why is choosing the right lab such a big deal? It’s actually something you CAN choose and it will make...difference for your future career and life. You might see a lab head as choosing you, but in reality, ...communication before making this important decision. I’ll highlight some of these questions in the sections below....you are good at asking for and getting help, size might not be a big factor for you. Know who's boss You...considering a career at a small, teaching college you might want to consider less expensive, portable areas ...ownership" and author order PI lets lab members fight it out PI intervenes if there are interpersonal...

  9. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    Published
    June 8, 2017, 2:30 p.m.
    ...excitation light less efficiently. Similarly, decreasing the power of the excitation light reduces the...series described a practical approach to selecting a bright fluorescent protein. In the second post of this...destruction of a fluorophore under the influence of light. Any fluorescent molecule will photobleach at some...fluorescent proteins are excited with very intense light (for a brief time), whereas in widefield imaging...imaging fluorophores are excited with relatively low light levels (for a prolonged time). Other illumination...that, in experiments that measure photostability, high excitation powers are often used to reduce the length...linearly related to power, the conclusions reached at high power may not translate to real applications in ...

  10. Zebrafish as a Model for Behavior: Swimming into the Optogenetic Spotlight

    Type
    Blog Post
    Published
    July 16, 2019, 12:59 p.m.
    ... short generation times and high fecundity, make zebrafish ideal for high throughput studies of optogenetics...Optogenetics, the use of light sensitive proteins (opsins) to manipulate cell activity, enables researchers... studies because it is non-invasive and offers a high degree of control over time and space. Zebrafish...imaging with fluorescent tags and activation of light sensing proteins. Furthermore, during the first ...Optogenetics functions through opsins which are light sensitive proteins. Opsins used in optogenetic studies...particles to cross the cell membrane when exposed to light. Neuronal firing depends on these charged particles...of the differing peak absorption wavelengths and light intensity requirements of different opsins, studies...

  11. Tetbow: Bright Multicolor Labeling for Neuronal Tracing

    Type
    Blog Post
    Published
    Jan. 24, 2019, 2:24 p.m.
    ...solution for discriminating between neurons for light microscopy-based neuronal reconstruction. To achieve...et al., 2007; Cai et al., 2013). However, the brightness of Brainbow was inherently low. This is because...at ~2 copies per cell per color to achieve the highest color variations. Adjust the plasmid concentrations... to express a minimal amount of tTA to achieve highest expression levels. Clearing large tissues - Fluorescent...anatomical aspects of sensory circuit development using light microscopy. Lab website: https://dn-en.med.kyushu-u.ac.jp...clearing and index matching of tissue samples for High-resolution fluorescence imaging using SeeDB2." Bio-Protocol...Sakaguchi, Richi, Marcus N. Leiwe, and Takeshi Imai. "Bright multicolor labeling of neuronal circuits with fluorescent...

  12. Enabling high school research at the Journal of Emerging Investigators

    Type
    Blog Post
    Published
    Sept. 5, 2018, 12:01 p.m.
    ...Investigators, an open-access journal that enables high school students to publish peer-reviewed scientific...behind the journal and hear from a fantastic team of high school students who recently submitted to the journal...Journal of Emerging Investigators. Chapter 2 - Getting high school students and teachers interested in JEI (...mentorship programs. Chapter 6 and Outro - Interview with high school student researchers Arooba Ahmed, Jiachen...

  13. Choosing the B(right)est Fluorescent Protein: Aggregation Tendency

    Type
    Blog Post
    Published
    June 15, 2017, 2:30 p.m.
    ...separate proteins based on size. Whether higher molecular weight complexes are detected will depend on the...series described a practical approach to selecting a bright fluorescent protein and a photostable fluorescent... have a natural affinity and a tendency to form higher order aggregates. This property can be tolerated...purified fluorescent proteins to form homodimers or higher order oligomers can be analyzed by several techniques...essential to verify a number of key properties (brightness, photostability, oligomerization) of a number...will be using. If you do a head-to-head comparison right, it will be valuable information for the scientific...

  14. A Practical Approach to Choosing the B(right)est Fluorescent Protein

    Type
    Blog Post
    Published
    June 1, 2017, 2:30 p.m.
    ...molecule. A higher brightness increases the detected signal and therefore high brightness is an important...EGFP). The higher the number, the higher the theoretical brightness. The theoretical brightness often appears...are (i) brightness, (ii) photostability and (iii) aggregation tendency. Brightness Brightness refers ...practical brightness is a better criterion than theoretical brightness for selecting the brightest fluorescent...and the sample. Practical brightness A protein with a high intrinsic brightness that does not fold well ...how the brightness is best defined or determined. Here, we will explain how practical brightness is determined...protein. Theoretical brightness Two key properties of fluorophores that determine brightness are the extent ...

  15. Channelrhodopsins with improved light sensitivity for minimally-invasive optogenetics

    Type
    Blog Post
    Published
    Nov. 3, 2020, 2:15 p.m.
    ...with the flick of a light switch. Neurons are not typically persuaded to fire when light is shined on them...expression of light-gated ion channels such as channelrhodopsins (ChRs) makes them light-responsive. When...mm3). Current channelrhodopsins also need high-intensity light for activation which requires implantation... no blue light, the channelrhodopsin remains inactive and neurons don't fire. With blue light, ions flow...are highly constrained and poorly understood. It’s hard to design new channelrhodopsins with high-throughput...requires higher expression levels in order to activate a neuron. ChRgers, however, have a higher conductance...the light was turned off.  Together these two experiments determined that ChRger2’s improved light sensitivity...

  16. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    Published
    Sept. 3, 2020, 12:15 p.m.
    ...dark state and highest fold induction at 31X. By using a slit pattern during blue light exposure (470nm...proteins N713-pMag and nMagHigh1-C714, this new system had both low background and high fold-induction of ...reversibly activated by turning blue light on and off. As one might expect from the modular nature of Cas9...fused with high-affinity interacting proteins Coh2 and DocS, respectively. The use of far-red light is of ...particular interest since blue light has limited applications in vivo. UV and blue light is readily absorbed and...tissues. Additionally, high intensity exposure can cause DNA damage. Far-red light, however, can penetrate...protein. Stop light! Photoactive anti-CRISPRs halt Cas9 activity Anti-CRISPR proteins are a highly diverse ...

  17. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi

    Type
    Blog Post
    Published
    April 25, 2019, 3:01 p.m.
    ...mutagenesis for the brightest teal fluorescent proteins that were photostable and had a high 480/530 nm emission...mWasabi, a bright, monomeric green fluorescent protein. In vitro, mWasabi is 1.6-fold brighter than EGFP...-type termini. mNeonGreen was reported as the brightest monomeric green or yellow fluorescent protein ...protein at the time. It is 1.5 to 3 times brighter than the most commonly used GFPs and YFPs. Its excitation...changes in pH. Find mTFP1 plasmids here! mWasabi: a bright green fluorescent protein The team that developed...Hui-wang, et al. "Directed evolution of a monomeric, bright and photostable version of Clavularia cyan fluorescent...PMCID: PMC2292683. Shaner, Nathan C., et al. "A bright monomeric green fluorescent protein derived from...

  18. High Complexity Golden Gate Assembly with NEB’s DAD

    Type
    Blog Post
    Published
    Aug. 1, 2023, 1:15 p.m.
    ...ever used Golden Gate Assembly for cloning, you might be familiar with the rules of thumb for designing...way to design GGA overhang sets, particularly for high-complexity reactions? The researchers over at New...otherwise! They suspected a data-driven approach might allow for an increase in the complexity. In 2018...need to be strictly followed in order to generate high-fidelity reactions. (Need a refresher? The five ...100% GC.) In fact, the NEB® team’s data showed that high-fidelity reactions could be done using overhang ...sets that broke rules 3—5. Fidelity, it turns out, might depend more on the individual sequence(s) than on...build out three tools to help researchers design high-fidelity Golden Gate Assemblies and overhang sets...

  19. ReFigure: Save Scientific Figures into Dashboards and Share Your Insights

    Type
    Blog Post
    Published
    Dec. 13, 2017, 3:20 p.m.
    ...researchers, we wondered how we could make the insights gained during this time more visible and thereby... ReFigure saves time, knowledge and makes your insights discoverable. Watch this quick video to learn ... ground again and again as they don’t save the insights they glean from a reading session. To remember...hunting through a whole folder of papers to find the right experiment! The ReFigure extension also tells readers...ReFigure making it easy for others to discover your insights on a topic. Beyond collating other’s data, you...include your own data in a ReFigure. For example, you might include a confirmatory image to show that a previously...previously used reagent performs well, or you might publish a replication experiment that you conducted ...

  20. Cancer, Inflammation and Immunity - Harnessing the Body’s Defenses to Fight Cancer

    Type
    Blog Post
    Published
    July 6, 2017, 1:59 p.m.
    ...increasingly able to harness the body’s immune system to fight progressing cancers. It was inspiring to participate...from all fronts!”  Here I aim to compile a few highlights from the conference which reflect how cancer/...targeted engineering of the TCR locus. He also highlighted the UCSF Immunoprofiler - a human biopsy analysis...

  21. Meet an Addgenie: Insights into Grad School, Career Paths, and More

    Type
    Blog Post
    Published
    June 25, 2020, 1:15 p.m.
    ...hand, there are reasons why grad school might not be the right choice. I spoke with our Financial Operations...here. You also don’t have to go back to school right away. Shannon Rinaldi, Addgene’s Lab Manager, was.... I thought, ‘Let me pursue this, let me make a slight change so I can have flexibility and be up to date...biggest takeaways from my co-op. It has been so insightful to learn about different career paths, successes...

  22. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

    Type
    Blog Post
    Published
    Feb. 7, 2019, 2:16 p.m.
    ...range between 0 and 100) are then converted to a weight (which ranges between 0 and 1) so that they can...ranging from -40 bp to 40 bp) are also converted to a weight (which ranges between 0 and 1) (shown in the graph...Conclusion CrispyCrunch is now publicly-available for high-throughput design and analysis of CRISPR experiments...

  23. Crowdfight, a Platform to Boost Scientific Collaboration During COVID-19 and Beyond

    Type
    Blog Post
    Published
    June 1, 2021, 1:15 p.m.
    ...oneself that prompted us to create Crowdfight.   Figure 1: Crowdfight is a community of scientists who...making science.   How we started Crowdfight-COVID19 Crowdfight was born as a collaborative platform...collaborator for a specific task, Crowdfight will: 1) Find an expert with the right skill; 2) Confirm the expert...Ignacio Amigo and María Hernández Sánchez from Crowdfight. In March 2020, while most of the world was locked...did not have access to a P3 laboratory (BSL-3), a high-security facility needed to perform the experiments... them heard about our online platform, called Crowdfight, and filed a request for help. In just a few ... a research group in Scotland, who had just the right infrastructure and offered to collaborate with them...

  24. Plasmids 101: 5 factors to help you choose the right cloning method

    Type
    Blog Post
    Published
    Aug. 21, 2018, 12:31 p.m.
    ...the challenge of trying to figure out what system might work best for your particular purposes. We’re here...than others. In today’s post, we will focus on a high-level comparison of popular cloning methodologies...results. If you’re in a hurry, Golden Gate cloning might be for you. This methodology relies on versatile... in as little as 30 minutes! Alternatively, you might consider Gibson cloning. In this technique, you ...fragments in an order determined by their homology. You might also consider FastCloning, first described by Chaokun...approach should you use if the budget is particularly tight? Most cloning processes will require a PCR reaction... job done, provided that you’ve already got the right enzymes laying around the lab. If not, you’ll be...

  26. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    Published
    April 9, 2019, 1:13 p.m.
    ...engineering of fusion proteins is relatively straightforward. However, generating a fusion while keeping...discuss strategies to generate fusion proteins and highlight some aspects of their design.  Protein size and...situation Next to size, potential homodimerization and higher order oligomerization of fluorescent proteins is... the fluorescent tag should operate as an inert light emitting module, any tendency of GFP to homodimerize...fluorescent protein characteristics such as the brightness, photostability, maturation time, and acid sensitivity...termini of your protein of interest The most straightforward fusion protein is made by connecting the N-...the fluorescent protein-POI fusions, we generate straight fusions, without any linker. When both orientations...

  27. Opto-Nanobodies: Using Light to Manipulate Cell Signaling and Protein Purification

    Type
    Blog Post
    Published
    Nov. 19, 2019, 2:08 p.m.
    ...and wrap the column with blue LED lights. When you switch off the lights, OptoNBs bind YFP and non-specific... the blue lights. Which option do you choose?  Option #2 may sound like fiction, but light-controlled...dark-induced dark-induced Loop 6 blue-light induced dark-induced blue-light induced Table 1: Summary of the... effect of AsLOV2 insertion site on light-induced binding.  Light controlled protein purification with...while cycling between dark and blue light which revealed a blue light-dependent shift in mCherry fluorescent...cells: light-based control of signaling pathways. This is particularly exciting because using light to control...binding of their target protein is altered by blue light illumination. This ability to reversibly bind and...

  28. A New Optogenetic Tool Based on AraC Controls Gene Expression with Blue Light

    Type
    Blog Post
    Published
    June 17, 2021, 1:17 p.m.
    ... (blue light-inducible AraC dimers in E. coli) that turns expression on and off with blue light.   Inspired...domain with VVD, a light-responsive domain that dimerizes upon exposure to blue light (Fig. 1). Another...VenturaIf you follow Barbara Di Ventura on Twitter, you might have seen the video of her singing about the paper...Another blue light-inducible dimerization domain, Vaucheria frigida aureochrome1 (VfAu1), led to functional...most applications.   Figure 1: With blue light, the VVD domains of the BLADE dimerize and activates...under the pBAD promoter, can be controlled with light without the need of any cloning.     What advantages...BLADE activates transcription with the flick of a light switch and shuts off just as quickly. The original...

  29. Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity

    Type
    Blog Post
    Published
    Oct. 11, 2018, 12:30 p.m.
    ... focused on three different levels of assembly: High efficiency and accurate single-insert assembly Intermediate...destination plasmid. Antibiotic selection allowed for high throughput cloning efficiency measurements. We performed...confirmed the insertion of the Lambda insert at the same high frequencies as the antibiotic selection markers....lacZ cassette assembly was easily achievable with high levels of transformants and low backgrounds – so...Using these 4 bp overhang observations, accurate “high-fidelity” junction sets for both the 12- and 24-...complex assemblies were performed. It was found that high efficiencies and accurate assembly levels were indeed...in the number of transformants while maintaining high levels of correct assemblies) translate to faster...

  30. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

    Type
    Blog Post
    Published
    Oct. 17, 2019, 12:28 p.m.
    ...This is where FLEx enters the scene. FLEx: A light when night is about you! The FLEx (for flip-excision)...optogenetics? Optogenetics: Controlling neurons with light In 2005, a team from Stanford University, led by...ago with the discovery of microbial rhodopsins, light-sensitive proteins that absorb photons for energy...maintain membrane potential in response to visible light (Zhang et al., 2011). What do microbial rhodopsins...2000). Most neurons in the brain are not naturally light-sensitive, so the selective expression of opsin ...Optogenetic neuromodulation is made possible by the light: when hit with the correct wavelength, microbial..., 2011; Mei and Zhang, 2012). For example, blue light activates channelrhodopsins that, in turn, trigger...

  32. Walking beside Academia and giving it a high five: My transition from graduate work to a position at Addgene

    Type
    Blog Post
    Published
    July 2, 2015, 8:37 p.m.
    ...Scientist. I used to spend my days (and often my nights :D) engineering E. coli to produce biofuels in ... screaming from the lab like the cartoon on the right?!? I would love to tell such a dramatic story, but...This may come as somewhat of surprise to many of my high school friends. In my teenage years I was super ...can involve walking alongside it and giving it a high five. While screaming can be therapeutic and I’ve...

  33. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    Published
    May 5, 2020, 1:15 p.m.
    ...coronavirus using CRISPR-based SHERLOCK (Specific ​H​igh ​s​ensitivity ​E​nzymatic ​R​eporter un​LOCK​ing)... under either no excitation light, UV light or LED blue light.  Highlights: This is a one-pot, one-temperature...heat block, LED blue light illuminator or UV light illuminator. Approach: Highly similar to the SHERLOCK...captured by an antibody further along on the strip. Highlights: Of the 83 known COVID-positive samples tested... extraction but that may work only if there is a high enough (105 copies per microliter) viral load in...lateral-flow readout slip that allows rapid detection.  Highlights: Cas13a is not catalytically active in the presence... duplex and will not give a positive result for highly similar viruses. Unlike Cas12a, Cas13a does not...

  34. Which Fluorescence Microscopy Technique is Best for Me?

    Type
    Blog Post
    Published
    Oct. 10, 2017, 1:57 p.m.
    ...limit of a light microscope. Due to the wave nature of light, an infinitely small point of light will blur... to provide high 3D spatial resolution while also avoiding the toxic effects of high light doses to the...fluorescence microscopy provides a high contrast image in which a bright signal overlays a perfectly black...placed to only allow light from the focal plane to reach the detector and all light from above or below...for example, red light instead of blue to excite a molecule of GFP. Because red light has approximately...approximately half the energy of blue light, two photons of red light are needed to excite GFP compared to only ...objective can a a high enough density of red-photons be establish for this to happen.Using red light has a second...

  35. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

    Type
    Blog Post
    Published
    April 25, 2017, 2:30 p.m.
    ...UV-Violet Dark/Green N/A (high) High 503/ 518 Padron Monomer Blue Dark/ Green N/A (high) Low 503/ 518 rsCherry...specific wavelength of light. PA-FPs can be activated from low fluorescent states to high fluorescence states...fluorescence, to a brighter fluorescent state by exposure to a specific wavelength of light. The switch or...fluorescence when illuminated with 488nm light. However, when UV-violet light is presented first, it causes irreversible...illuminated with violet light, both variants are photoconverted to forms that emit brighter red fluorescence....when exposed to blue light, which is less phototoxic to live tissue than the UV light used by other PA-FPs...State Activating Light Pre/ Post Color Contrast (Fold Change in Fluorescence) Brightness Excitation/ Emission...

  36. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    Published
    June 27, 2023, 1:15 p.m.
    ...with exquisite precision using only light. Optogenetics, or light activated tools, can control everything...an experimental optogenetic setup. A light source delivers light pulses to the subject/cells expressing... photoswitches. Light-gated ion channels directly open and close when exposed to light and change the ...changes with light stimulation to either activate or inhibit a specific receptor or pathway.  Light-gated ion...turn on with a light-pulse at one wavelength and stay on until a second pulse of light, with a different...pathways with light. This is achieved by coupling endogenous signaling molecules with light-activated partners... intensities of light sources used to prevent activation of both tools with each light source used.   ...

  37. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    Published
    May 28, 2024, 1:45 p.m.
    ...dim" you might get the impression of a candlelight vs. a floodlight. For fluorophore brightness levels,...flow-cytometry-fluochrome-brightness-spillover Excitation Laser Color Very Bright Bright Moderate Dim...Photons of lower wavelengths are higher in energy, while photons of higher wavelengths are lower in energy...important to know the brightness of individual fluorophores. Table 1 describes the brightness of a few commonly...the cytometer. Brighter fluorophores have a stronger signal. Table 1: Degrees of brightness for a few commonly...-Cy7 are "very bright" when they are excited by the yellow/green laser but only "bright" for the blue ...leading to different levels of brightness, depending on the laser light's wavelength. Source: https://www.bdbiosciences.com...

  38. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    Published
    March 21, 2024, 1:15 p.m.
    ... to their brightness and high degree of overlap. Dyes for DNA/RNA staining should have high affinity and...refers to the emission of light by a substance (fluorophore) that has absorbed light or electromagnetic radiation...whereby excitation of blue light will indicate where FITC is bound through green light emission. Key considerations...also vary in their brightness, depending on the respective fluorochrome. Brightness is impacted by both...of findings. If you’re struggling to select the right fluorescent tools, this blog is here to help. Here...ligand to make your selection easier, as well as highlight some recent advancements within this field. Fundamental...wavelength (Heinrichs, 2009). When a fluorophore absorbs light, electrons are elevated from a ground state to an...

  39. Synthetic Photobiology: Optogenetics for E. coli

    Type
    Blog Post
    Published
    Sept. 8, 2015, 2:30 p.m.
    ...neuroscience. In fact, precise light-based control of biological processes is highly useful in other fields, ...consistent, and controllable pulses of light. Complicated patterns of light intensity and wavelength can be ...to low-intensity light, so off-target effects should be limited. Phytochromes are light-responsive systems...harness the power of light, Christopher Voigt’s lab created the first E. coli light-sensitive two-component...construct senses light, and an obligate chromophore allows the system to respond to various light inputs. In...various light inputs, and an obligate chromophore allows the system to respond. In far-red light or dark...exposure to red light rapidly deactivates the system. Version 1 of this far-red/red light-responsive system...

  40. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ...LACE system for light-activated CRISPR-Cas9 effector, Polstein and Gersbach fused the light-inducible protein...binding affinity with light stimulation2. The Kuhlman lab created two improved Light Inducible Dimers (iLID...of Arch mutants screening for improved brightness. The brightest mutants were then further mutagenized ...enhanced sdChr construct, named CheRiff, exhibits high light sensitivity and fast kinetics. By combining both... weak or high activities (Bert, et al. 2000). To design pXPG, the authors incorporated a high-copy origin...repository, we can't give all of them attention they righlty deserve, but, in this post, we'll provide a small...2015 Feb 2;33(2):139-42. doi: 10.1038/nbt.3149. Light-inducible CRISPR-Cas9 system for endogenous gene...
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