Cytometry and Antibody Technology

Introducing the Agilent Penteon 5-30

by | Mar 11, 2022 | Announcements, Instruments | 0 comments

Flow cytometry equipment has improved quite a bit over the last few years and the CAT Facility aims at offering the best instruments available on the market. In the aftermath of the Flood of ’21, we ended up replacing the irremediably damaged Fortessa X20 by a Penteon equipped with 5 lasers and capable of picking up up to 30 fluorescent parameters. There’s a number of features that we think are going to be incredibly appreciated by our users, and we will share them here.

Let clear one thing first however: Can the instrument really pick up 30 markers? Well, it has 30 detectors, but some of them are likely not going to be usedall that much. Just on the blue line, we have a series of detectors for PerCP, PerCP-Cy5.5, and PerCP-Alexa710. That’s more about being able to taste the right flavor of PerCPs than getting really useful channels. So what’s going on? Simply put, the cells are hit by 5 spatially separated lasers and the signals are going through the same set of dichroic mirrors and bandpass filters to get to their specific detectors. Still, with all the detectors available, you’ll be able to use the vast majority of fluorophores available on the market, and in fairly complex panels.

Emission and filter set for PerCP, PerCP-Cy5.5, and PerCP-eFluor 710. There is too much overlap between all three fluorophores and using them in the same panel would be problematic.

Speaking of detectors, the Penteon uses silicon photomultipliers. Check out Bert’s post PMT vs SiPM: A Photon Finish for a reminder of how they work. One key feature of these bad boys is that they have a high dynamic range which reduces the chance of them getting saturated. Very bright fluorescent proteins might do the job, but regular antibody staining are otherwise unlikely to blind the detectors at the default gain settings set by the manufacturer. Users will have a much lower chance of losing an experiment due to poor gain decision, and the default gains will usually work fine for most applications. Would you want to tweak them for your own purpose? Sure, be certain that the cornerstone figure of you future publication will look as good as possible. But the data will likely look pretty nice from the get-go.

Automation is another feature that we really enjoy about the Penteon. There is something in me that dies a little every time I see a users feeding tubes to a Flow Cytometer for hours on end. The NovoSampler Q is a reliable autosampler that will handle plates and tubes alike. It will vortex and rinse the probe according to your specifications. A soon to be purchased adapter also allows your samples to remain at 4C for those long bouts of acquisitions.

As a core manager, what really seals the deal for me is the ease of maintenance of the instrument. A simple 3 tube protocol has been set up to clean the probe after each experiment, and the instrument has its own cleaning protocol that handles the rest of the fluidic line and the flow cell every time it is powered down. Furthermore, the Penteon does not allow users to run the instrument while the sheath tank is empty, or any other self-destruct features. The users are no longer at the mercy of the previous individual who forgot to something or another about proper care of a Flow Cytometer. This should save us all a lot of heartaches in lost experiments.

Special mention to the automated compensation wizard. The auto-gating is spot-on most of the time!

By the way, we also offer a Quanteon, the Penteon’s smaller sibling. That instrument has 4 lasers, can pick up 16 parameters, and is located in the the Jonas Center in room S301 of the Billings Hospital. The NovoExpress software is intuitive and most of our early adopters lose no time figuring out how to get what they need. A offline version of the software are available on analysis stations of both the main lab and the Billings satellite facility. So have a look at the Penteon’s page, give the instrument a shot and let us know what you think!

Here is some training material for the Novocyte platform!

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