Fast analysis of acetylcholine for neuroscience applications

Fast analysis of acetylcholine for neuroscience applications

Fast analysis of acetylcholine for neuroscience applications

We show you how to detect and analyze acetylcholine in brain microdialysis samples in 18 minutes, with minimal or no need to add an acetylcholine esterase inhibitor or AEI.

For this analysis, we use high-performance liquid chromatography or HPLC, coupled with an immobilized enzymatic reactor column and an electrochemical detector. It’s fast and highly sensitive, down to the femtomole range.

Acetylcholine is separated by a polymer-based reverse phase column, which has been selected specifically for this application. Not only is the column we’ve selected great at separating acetylcholine, but it can also handle the higher pH required for optimal enzyme activity. After exiting the separation column, acetylcholine and other compounds enter an immobilized-enzyme reactor column. Inside the reactor column, compounds are broken down to produce hydrogen peroxide, which is then selectively oxidized by an applied voltage across the flow path and changes to current are detected with a platinum electrode.

Optimizing the right conditions can take time and resources. That’s why many labs require highly-trained specialists in HPLC. However, we’ve created an application, which includes everything you need to run the acetylcholine analysis right away.

For this application, we’re going to use the HTEC, which includes everything you need integrated into one single unit. Let’s take a look inside:

-Dual-piston pump with a unique algorithm to reduce noise without any pulse damper.
-Degasser to remove small air bubbles for better pump performance
-Temperature control for consistent results
-Separation column uniquely selected for acetylcholine
-Enzymatic reactor column
-Electrochemical detector cell with a platinum working electrode for selective detection of the enzymatic reaction products.

We’ve optimized conditions to make it as easy as possible. Simply inject the sample using a manual injector or autosampler if you’re using laboratory automation. You can monitor signal response in real-time using the dedicated chromatography software. In 18 minutes you will have your results and you’re ready for the next sample. It’s highly sensitive down to the femtomole range. Here’s an example of a chromatogram showing acetylcholine.

For more information on acetylcholine analysis, contact us today!


Precise Touch Operant Training

Precise Touch Operant Training

Precise Touch Operant Training

Today I’m going to demonstrate the unique features of our touch panel operant chamber system.

What is unique about our chambers is that they contain infrared sensors located at the top of the touchscreen itself that improve the accuracy of touch responses from small rodents.

Unlike the touch screen technology that most of our smartphones and computer screens use, called projective capacitance, the infrared sensors inside the touch screen improve accuracy by eliminating the need for a minimum force required to generate a response. This means that even nose poke’s from mice will register a response.

In addition to improved sensitivity, our touch panel was designed in a trapezoidal shape instead of a square, making it easier for the animal to focus on the screen ahead.

Our touch panel chambers are compatible with in vivo electrophysiology and optogenetics techniques, as well as with miniature head-mounted microscopes.

Chambers can be purchased singly or in a package of four for a more cost-effective option. In addition, the chambers come with our Operant TaskStudio software package, an extremely user-friendly software platform that enables customers to design and execute their own tasks or choose from a variety of pre-programmed tasks.

Check out more about our software in our next video. To learn more about our Touch Panel operant system please check out our product website or connect with an expert.


Benefits of using HPLC-ECD for Neurotransmitter Detection

Benefits of using HPLC-ECD for Neurotransmitter Detection

Benefits using HPLC-ECD for neurotransmitter detection

Here’s why we recommend HPLC-ECD for neurotransmitter detection.

While there are a few ways to analyze neurotransmitters, like liquid chromatography-mass spectrometry, or LC-MS, ELISA, or Radioimmunoassays. However, for qualitative and quantitative analysis of neurotransmitters, we recommend high-performance liquid chromatography detected with an electrochemical detector, known as HPLC-ECD.

The first reason is sensitivity. HPLC-ECD is highly sensitive, down to the femtomolar range. This is suitable for most neuroscience applications, including a quantitative analysis of in-vivo microdialysis samples.

The second reason is time. Applications using HPLC-ECD can run anywhere from 5 to 30 minutes, where you can detect multiple analytes from a single injection. For example, we have an application for the simultaneous detection of dopamine and serotonin in just 5 minutes. That means you can run over a hundred samples overnight for high throughput using laboratory automation like an autosampler. We’ve also developed applications, saving you the economy of time by already optimizing conditions. All the applications on our site we know work, because we have tested them in-house.

The third reason is price. HPLC-ECD is priced with intended use in your own lab, thus, no need to outsource analysis in another lab or core facility. This gives you, the user, more confidence in your data knowing that you are in control. Pricing for the equipment typically ranges from $20,000 to $60,000. If you’re not quite ready to commit to a new machine, ask us about our leasing options.

Finally, usability. While HPLC can seem overwhelming and complicated, we’ve simplified it with the HTEC. The HTEC system is an all-in-one device that integrates everything you need into one instrument. It’s straightforward and easy to use. Should you decide to acquire one of our instruments, on-site installation and training at your lab will be provided. Once installed, you can immediately start running samples. If you have questions or need assistance, there’s no need to worry. Our self-help support site and knowledgeable support staff are available to guide you through the process, including how to use it as well as help with troubleshooting. At Amuza, it’s important for us that our instruments work just as well for you in your hands as they do for us.

Therefore, we recommend HPLC-ECD for the detection of neurotransmitters.
For more information, 
contact us today.

How the ENO-30 Detects Nitrate and Nitrite

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An overview of the ENO-30, which can simultaneously detect nitrate and nitrite in biological and other samples.

The ENO-30 is a uniquely designed system that combines HPLC with a colorimetric assay optimized for fast and highly sensitive quantitative analysis of nitrate and nitrite. By utilizing the ENO-30, you can get accurate data in 10 minutes from a single injection. Combined with laboratory automation using an autosampler, you can run hundreds of samples overnight.

Let’s take a look inside to see how it works: The ENO-30 has an upper and lower unit. The lower unit contains the HPLC pumps optimized at just the right pumping speeds for the proper mixing of reagents.

Samples are injected with either a manual injector as shown or with an autosampler. Both nitrate and nitrite are first separated by retention time with an HPLC column. After separation, nitrate is reduced to nitrite. The system utilizes the Griess reaction, in which each nitrite forms an azo dye and is detected as the pink color develops and is measured by absorbance detection at 540 nm.

The design of the entire system has been optimized for highly reproducible results. It’s simple and easy to use. Here at Amuza, it’s important that instruments we provide work just as well for any given end-user as they do for us here at our own facility. That’s why we offer installation and training, as well as product support for all our products.

Thank you for watching. Feel free to contact us if you would like to learn more about running nitrate and nitrite samples.


Obtain accurate fluid intake measurements

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The Drinko Measurer is one of our most simple products, but an extremely useful one for gathering precise measurements of liquid consumption in animal studies.

What makes the Drinko Measurer unique is that it contains double ball bearings inside the nozzle of the sipper, preventing leakage of excess liquid. Furthermore, when force from a mouse or rat is applied to the tip of the sipper tube, a single drop of solution is applied.

Each tube also contains a clip that attaches to the sipper tube to the cage to stabilize the bottle to the cage. This prevents it from being knocked out of place by the mouse or rat. This clip can also be adjusted so you can adjust the length of the nozzle within the cage.

Additionally, all parts are autoclavable for easy cleaning.

The Drinko Measurer comes with several different sipper tube lengths 2.5, 3.5 and 4.5 inch as well as two different bottle sizes, 10 and 15ml. We also have new 15ml bottles with ml markings making it easier to visualize liquid consumption. However, we recommend weighing the bottles for the best results.

Now I’m going to tell you about several applications we recommend for the Drinko Measurer. The first is for drug delivery experiments.
For drugs that can be administered orally and easily mixed into a solvent, simply use the Drinko Measurer as a means for drug delivery rather than having to inject your experimental animals.
This is useful for drug dosing experiments and toxicology studies. Also, by combining two units per cage, you can measure drug-seeking preference with drugs of abuse.

The second application is for conditioned taste preference. Or conditioned taste aversion
Easily Measure preference or avoidance of a liquid by combining two Drinko Measurer systems and measuring liquid consumption.

Finally, we do carry 40 mL bottles which are useful for larger animals or for extended access studies. 

You can purchase units easily by navigating to our shop.


Factors that Influence Experimental Outcomes and How to Overcome Them

Factors that Influence Experimental Outcomes and How to Overcome Them

Experimental outcomes can be influenced by a variety of factors, some of which can be controlled for. Minimizing confounding factors is crucial to gathering reliable and repeatable results.

One of the biggest issues in animal research today is the replicability of results. Too often animal study outcomes can not be repeated. This is not hard to believe given that the use of animals themselves provides inherent variability, even when all other factors are controlled for. Differences in the strain of animals used, as well as the age of the animals, time of day that experimental tests are administered, and how long the animals were handled prior to experimental testing are just some of the factors that can impact experimental outcomes.

While it is impossible to eliminate all external factors, animal-experimenter interactions can have a huge impact on results and should, therefore, try to be minimized as much as possible.

How can you mitigate animal-experimenter interactions?

At AMUZA, we offer a variety of automated behavioral tests that were designed specifically to improve the reliability and repeatability of behavioral assays.

For example, our Touch Panel operant training system is an automated operant chamber that utilizes photo beam sensors in the touch panel itself to improve the accuracy of responses from small rodents. The Touch Panel also includes software that enables users to design and run their own tasks with video tracking capabilities for automated data collection.

Even our standard mazes come with video tracking and automated data collection and analysis.

Touch Panel

Self Head-Restraining Platform

Furthermore, one of our other products, the Self Head-Restraining Platform, was designed to completely automate the head-fixation process in mice in order to streamline head-fixed behavioral assays.

In fact, the platform, originally developed by Dr. Andrea Benucci at RIKEN brain institute, was designed specifically to help overcome the reproducibility crisis.

Not only do our tools free up experimenter time and labor to focus on the actual science, they help remove unwanted experimenter bias by standardizing the experimental testing arena.

Even with automated behavioral tasks, however, it is still possible to introduce experimenter bias. This is why we also recommend that you perform rodent behavioral tests at roughly the same time each day, as well as handle experimental animals equally. Ideally, the same experimenter should be handling the animals each day. If this is not realistic, different experimenters should be counterbalanced across days, or across testing groups.

Also, if you plan to use different strains of mice or rats for your experiments, make sure to run behavioral tests across these different strains to account for any strain-specific differences.

Additionally, with our automated rodent behavior systems, we recommend that the motivation of the experimental animals to perform the task is consistent. If animals are food or water-deprived, weights should be taken daily initially and then weekly thereafter to ensure that test subjects are maintained at similar percentages of their free-feeding body weight.

Cleaning the testing chambers between use

All of our behavioral tests are made out of acrylic that is easy to clean as well as removable floors. Testing arenas should always be cleaned between experimental sessions to make sure the scent of the previous animal will not influence behavioral results.

For more detailed information about our automated behavioral tests connect with us today.