... 2022). The oxytocin sensor MTRIAOT enabled real-time observations of oxytocin dynamics in live mice under...here!   Ino D, et al. A fluorescent sensor for real-time measurement of extracellular oxytocin dynamics in...synaptic glutamate response. They generated variants optimized for different applications (v82 has higher affinity...indicator (GEVI) called JEDI-2P (Liu et al. 2022) optimized for two-photon imaging. They use a unique and ...from Gary Yellen’s lab is LiLac, a fluorescence lifetime sensor for lactate developed using a high-throughput... for performance under two-photon fluorescence lifetime imaging. B) LiLac responses are highly resistant...multiparameter screen for accelerated development and optimization of soluble genetically encoded fluorescent biosensors...

...non-invasive and offers a high degree of control over time and space. Zebrafish have become a popular model...reflex. These traits, coupled with short generation times and high fecundity, make zebrafish ideal for high...neuron more positive or more negative.  Neuronal stimulation: Depolarization When the inside of the neuron.... When startled by acoustic, visual or tactile stimuli, larval zebrafish react by performing a distinct...behavior is not done in response to any specific stimulus. While the escape response neural pathway is well...the overlap between groups.  They found that photostimulating the Kolmer-Agduhr cells was sufficient to ...Kolmer-Agduhr cells on one side of the body required photostimulation in order to drive the full symmetrical response...

...temperatures for long time periods and workflows where CRISPR nuclease activity is desired and time or protocol ...the hot springs My husband, Paul Blainey, at that time a post doc in the Quake Lab, volunteered to do the...joined for half of the trip. Preparations ahead of time included contacting YNP staff to arrange permits... the 3,471 square mile size of YNP, it took some time to map a route that would reach all of the remote...IgnaviCas9 protein is 1,241 amino acids long, has optimal sgRNA spacer length of 22-25 nts, and is thermostable... Cas9 from Parvibaculum lavamentivorans, whose optimal growth temperature is 30°C. Find IgnaviCas9 here...

... changes of these Twitch sensor variants were optimized by testing various linker configurations in a ...screen in rat hippocampal neuron cultures. The optimized Twitch sensors allowed the researchers to see ...he recommends they give it a shot. “Beginners sometimes think it’s complicated, but actually it is very...that the approach he and his colleagues took to optimize sensitivities can now be applied to dozens of ...simple swapping of binding domains, in practice optimizing the sensors involves plenty of trial and error...between different molecules like zinc and calcium. Ultimately, Merkx said he hopes others will find his tools...s like a test.” References Thestrup et al. Optimized ratiometric calcium sensors for functional in ...

...experienced mentor. The lab was very, very quiet. Every time I asked a question, I felt like I was yelling. I...information you will need is about you. It’s worth taking time to honestly evaluate your own style of work and ... help identify your priorities and needs, but sometimes it just takes introspective thought on what’s ... thrive. How big is the group? Has it grown over time? If not, why not? It may not be a bad thing if the...humiliation, accusations regarding lack of effort, intimidating use of discipline or competence procedures Threats..., persistent teasing, name calling, insults, intimidation, sexual harassment Isolation – including preventing...Do lab members work on more than one project at a time or over their stay in the lab? Do all lab members...

...platforms like HaloTag® Fusion Protein, codon-optimized Firefly luciferase genes (e.g., luc2), and NanoLuc...issues as traditional luciferases emit at near optimal wavelengths (Fluc emits at 610 nm) and do not require...’ dim light emissions requires long acquisition times that may be impractical or infeasible for certain... emission peak is at 460 nm thus making it non-optimal for in vivo imaging. The solution is to couple ...quantum yield, brightness and improved maturation time compared to similar orange fluorescent proteins....et al. (5) imaging multiple proteins at the same time in live cells using fluorescent proteins (FPs) can... variants of bright luminescent protein for real-time multicolour bioimaging. Nature Comm. 7, 13718. PubMed...

...nucleases are promiscuous. In other words, they can sometimes localize to unintended sites with sequence similarity... mutation burden. The CRISPR platform must be optimized to reduce off-targets and patient risk. Hopefully...experiment, at least one will likely be useful. Optimal gRNA selection With some CRISPR experiments, there...can reduce off-targets at a fundamental level. Optimal Cas selection Once you’ve selected a gRNA with ...elsewhere in the genome to reduce off-targets, it is time to consider your Cas enzyme. Even if there are similar... spCas9, has gone through multiple rounds of optimization which have led to more specific variants. HypaCas9...Rabinowitz and Offen). Of note, most of these optimized Cas enzymes prevent cutting at mismatched guide...

...where you met and what you discussed. Do this in a timely manner (within a few days or a week) so they can... to build relationships and strengthen them over time.  Things to avoid when following up with your ...connect via LinkedIn with strangers. It’s a waste of time. People that don’t know you, can’t really help you...people who do respond. Don’t call or email multiple times if you don’t get a response. It breaks an important...them and they don’t know me. It’s a waste of our time.  Networking connections can last years This is...ask me about the different arms of this starfish sometime, but as of yesterday, I have 4,700 LinkedIn connections...necessary. Once you’ve talked with a person a few times, if it was a positive experience for you both, they...

...working hours (9am-5pm Mon-Fri or overtime and weekends, part time or full time) and what life is like for someone...correctly the first time. These skills are very desirable to employers because they save time and money and ...attracted to science because they enjoy spending time outdoors and with nature. Experience in the field...assessed your skill set and done your research, it’s time to start sending out applications. Check out our...

...Years later, these classic systems continue to be optimized by Tabor’s lab, making light-controlled gene expression... biological processes. Chemical effectors are suboptimal for a number of reasons, including their potential...high or very low levels. Schmidl et al. chose to optimize these systems to make them more tunable and user-friendly...transcription factors, for which low expression levels are optimal. These tools are compatible with many strains of...et al. also represents a large step forward in optimizing TCSs. Over 75,000 TCSs have been identified in... in bacteria, and with similar optimization methods, many of these systems could be made suitable for ... applications.   References Refactoring and optimization of light-switchable Escherichia coli two-component...

... available amino acids a bit…limiting. It may be time, then, to look towards genetic code expansion (GCE... which non-canonical amino acids (ncAAs, also sometimes called unnatural amino acids) are placed in a ...along with the inherent trickiness of using and optimizing GCE, led to a large toolbox filled with systems...dubious enough that they don’t give it a try. Optimization Why is GCE so complicated? Well, while the techniques...growth temperature) may need to be individually optimized for each of the three factors: the ncAA, the tRNA... synthetases. And, naturally, rarely are the optimization parameters the same for all three components...including their phosphoserine ncAA protocol, to optimize and share, they’re soliciting feedback from scientists...

... a full-time career or a part-time passion, there are many ways to make it work for your time and interests...is science policy?”  I have been asked this many times throughout my short science policy career and this...including fellowships, volunteer positions, and part-time opportunities. But even through all my roles and...academia as tenured professors. There’s never a bad time to start your science policy journey.  In fact, ...Note: the answer to this question may change over time and that’s perfectly normal!  Action Items:  Start...presenting for non-scientist audiences during your time in graduate school through outreach organizations...

...this really sounds like a two-way ANOVA. And this time, it is! Factorial ANOVAs include two-way ANOVAs ...impact of temperature, humidity, sunlight exposure time, and watering volume on total crop production (though...the only things that matter. However, this is oftentimes an oversimplification. If your independent variables...every pairwise comparison within your population? Sometimes this can’t be avoided, but if it can be, you can...example, if your alpha is set at .05, then 5% of the time the statistical test will be incorrect. If you have...high false positives. Control vs. all samples Oftentimes there exists a control group and multiple experimental...working with, graphing data with a histogram is sometimes sufficient to determine normality (or lack thereof...

...that the optimal buffer for Taq DNA Ligase contains 100 – 200 mM KCl at pH 8.5. This optimization method...condition optimization, will be critical for success. Online tools are available to help optimize ligation... the incidence of annealed mismatched dsDNA. Optimization through high-throughput profiling New England... both the enzyme and the reaction buffer were optimized, resulting in the highest fidelity NAD+-dependent...Reaction Temperature Calculator can be used to estimate an appropriate incubation temperature when using...: 23842410. 19. LeClair, Norbert P., et al. "Optimization of a ligase detection reaction-fluorescent microsphere...

...combined optical and genetic methods to photo-stimulate mammalian neurons and alter synaptic events: a...rhodopsins and neurons have in common? Neuronal stimulations are triggered by the movement of ions across...viral-infected neurons are subsequently photo-stimulated through fiberoptic cannulas directly implanted.... In particular, they found that optogenetic stimulations of the DA neurons produce behavioral and electroencephalography...could activate appetitive rather than aversive stimuli. In a recent paper, de Jong and colleagues used...neurons of the VTA. Using in vivo optogenetic stimulations, they simultaneously recorded the electrical...PMC3225502. Boyden, Edward S., et al. "Millisecond-timescale, genetically targeted optical control of neural...

...Before diving into the experimental work, spend some time outlining the construction of the plasmid and all.... It is always a good idea to perform primer optimization, especially if you are having difficulty amplifying... genome, you can recheck your primer design or optimize your primers or PCR conditions. Purifying DNA...increase your chances to have a successful and ultimately faster cloning. Remember to quantify the DNA ... parameters that you can adjust: temperature and time of incubation, and amount of DNA. As a general rule...plasmid:insert), but this parameter has to be optimized according to the strategy adopted, and to the ...and if the strain is growing fine, it is probably time to prepare new plates with a fresh stock of antibiotic...

... do you have to work with difficult people? Each time I ask this question at my workshops, I get nods ...impolite, Complainers, who spend most of their time moaning about their troubles, Silent or unresponsive...supervisors: Hands-off super busy types, who never have time to mentor you, and Micromanagers, who question.... You can save yourself and your supervisor some time if you come to every meeting with one or more proposals...Your supervisor will appreciate that you took the time to think of solutions, and will be more likely to...your action item list and set some approximate timelines. Be sure that you keep your commitments, and if...your point across. Some supervisors might try to intimidate you with negative comments (hostile-aggressives...

...conversation to pass the time in a job that wasn’t going anywhere, but it became a legitimate brain exercise in...still flip to that page in my lab notebook from time to time. I wrote in large letters “Going for it!” We...there very few jobs available. So, we spent a lot of time talking about what we could do to make ourselves...weren’t restricted to science jobs. Keira’s an optimist, I’m a realist, and together we make a pretty ... 2014. Outside of the lab, Keira enjoys spending time with her husband and two Jack Russell Terriers. ...

... and significant lengths of time, the stability of fluorescence over time is crucial. For instance, the... century, it can be overwhelming to select the optimal dye or fluorescent ligand when designing an experiment...intensities, and the best practice to determine optimal DOL is through several small-batch experimental...can be mitigated through careful selection and optimization. Fluorescent experiment types: Different types...super-resolution microscopy techniques, such as Stimulated Emission Depletion (STED), and Stochastic Optical...non-fluorescent states in response to a specific stimuli, which leaves only a small volume of fluorophores...technically cell-permeable, its use in live cells can sometimes result in cellular toxicity due to the permeation...

...applications (qPCR and ddPCR.) qPCR Quantitative/real-time PCR (qPCR) has been used for copy number assays ...or negative for each reaction, math happens (this time, Poisson statistics), and the readout gives an absolute...Gommersall L, Arya N, Patel HR. Basic principles of real-time quantitative PCR. Expert Rev Mol Diagn. 2005 Mar...Quantitative analysis of copy number variants based on real-time LightCycler PCR. Curr Protoc Hum Genet. 2014;80:...hg0721s80 Colborn JM, Byrd BD, Koita OA, Krogstad DJ. Estimation of copy number using SYBR Green: confounding ...Na-Bangchang K. SYBR Green I and TaqMan quantitative real-time polymerase chain reaction methods for the determination...determined by SYBR-Green-based quantitative real-time PCR. J Microbiol Methods. 2006 Jun;65(3):476-87....