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Facility News
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Seeing as the bookings are plummeting, I can only assume that you either have all published every single paper you worked on this year or took the summer break ☀️ you deserve ! Either way, it is a good time to remind you that we are available to help you design panels, troubleshoot experiments and brainstorm your latest stroke of genius ! Just ask the FCF Staff !
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In this month FACS Tips, Kevin is giving us a recap of CYTO 2023, the biggest flow cytometry convention of the year.
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Silvia Arroz Madeira won the mug this month, Congratulations !
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Each month, we will give away one of those special and unique mug designed by the FCF team. Answer correctly to our 3 questions and you can have a chance to win !
Please take few minutes to answer the quiz HERE.
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FACS Tips
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CYTO 2023 Recap
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It was my privilege to travel to Montreal, Canada and attend CYTO this past May. This is the biggest annual conference in flow cytometry and the best chance to see what’s new and developing in the world of cytometry. As with last year, I will overview some of the new and interesting technologies presented at this year's conference.
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BD FACS Discover S8
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Image from https://www.bdbiosciences.com/en-ch/products/instruments/flow-cytometers/research-cell-sorters/bd-facsdiscover-s8
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Certainly, the highlight of this year's CYTO was a working presentation of BD’s new sorter. A first of its kind machine, capable of fluorescent imaging and sorting at high speeds. This machine presents massive opportunities for future research, and seems to be the leading edge of where machine technology is headed. Up to 3 fluorescent imaging channels off only the blue laser have been included on this machine, along with FSC, SSC, and light loss imaging, and these parameters can even be used to sort cells in real time. So, while this machine may not compete with the 12 imaging channels and 4 lasers of our ImageStream machine as far as imaging, it offers an interesting hybrid of speed and sorting capabilities that can’t be provided anywhere else currently. The Discover S8 is also the first to include BD’s new SpectralFX technology, which is its new spectral algorithm, utilizing its 78 detectors it has a massive range for panel size and specificity. One of the most exciting things about CYTO is getting to see and test new hardware, and get a sense for what the next big thing is, and this machine has certainly set a high bar for everyone else to match. We’re very excited to see what comes next.
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BD Horizon Reagents
To keep up with the expanding machine capabilities, BD has also released a new group of reagents under the name Horizon. The goal of these reagents is to expand the detection range for the Blue, Yellow, and Ultraviolet lasers respectively. These new dyes are designed to have lower amounts of cross excitation, while also being much brighter than some of the legacy dyes you would be used to in this emission space. For example, BD Horizon Brilliant Blue 515 (BB515) is claimed to be both brighter than FITC, while also producing lower spillover into PE. These dyes will certainly be an advantage for larger spectral panel development. And if the antibody you need isn’t currently commercially available, BD offers a new program called OptiBuild where you can custom order your chosen marker to a desired antibody. They advertise the possibility of over 24,000 different possible marker fluorophore combinations, and they can be produced quickly in small quantities. This can be massively helpful in adding markers to pre-existing panels to avoid losing both the time and financial investment to a complete panel redesign. Lastly, BD has also produced a complimentary Horizon Brilliant Stain Buffer to get the most out of these new reagents by ensuring reduced interactions between polymers.
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CYTEK Cloud and Synchronization
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Image from cloud.cytekbio.com
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Good news for all our Aurora users (and potential future users)! With CYTEK’s new Cloud digital ecosystem you can get up and running faster on the aurora. This service offers three tools; Full Spectrum Viewer, Panel Builder, and Experiment Builder. The Full Spectrum Viewer allows you to see and compare the various fluorophore spectrums to have an idea what will and won’t work together, or where the peak excitation of a fluorophore is. The Panel Builder will help you to design a spectral panel, and optimize to reduce spillover and complexity, and includes a spectral viewer at the end. While this tool doesn’t appear to be as robust as the EasyPanel software for panel development, there may be features you find preferential. Lastly there is the Experiment Builder tool, which allows you to set up the template of your experiment before you get on the machine, saving you a lot of time, and money!! From tube names, label names, fluorophores, and acquisition settings, these can all be chosen beforehand and then just loaded into the SpectraFlow software when you arrive at the machine. (https://cloud.cytekbio.com/)
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Ghost Cytometry
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Undoubtedly the coolest sounding technology at CYTO, ghost cytometry is a technology that allows you to sort cells based on an image, but without ever producing an image, or potentially even without fluorophore labeling. While certainly a complex technology to explain succinctly, ghost cytometry relies on collecting the compressed spatial information required to produce an image, but due to the constraints of processing speed required for sorting it processes these measurements without producing the image using AI, such that sorting becomes possible. It looks at cells, not as a focused excitation point, but rather spread out across a meshed screen, which they call a structured illumination. This generates almost like fingerprints for the different categories of cells, which can then be selected and sorted. Two of the groups in this space at the moment are ThinkCyte with its VisionSort (https://thinkcyte.com/), and DeepCell with its REM-I sorter (https://deepcell.com/).
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Sample Prep Technologies
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With machines capable of detecting over 50 to 60 parameters, and companies producing reagents to fill the necessary fluorophore gaps to produce the unique spectrums to make up those parameters, the next bottleneck in the workflow becomes the ability to reliably stain those large antibody panels in many samples. Even with small panels we can make mistakes keeping track of what antibody went where during staining, now imagine 12 colours turns to 50. Not to mention the time and energy it takes to handle that amount of organization and preparation. Multiple companies have begun to develop auto processing machines. CYTEK has developed the Orion Smart Cocktail Reagent Mixer, which can automatically prepare your master mix for you. With a programmable interface, it can hold up to 60 reagents and prepare your custom antibody mixes quickly and consistently. A more all in one solution has been developed by Curiox with their Laminar Wash Auto 1000 System, which can wash, stain, and even fix and perm your samples automatically for you. This can save massive amounts of time. Their Laminar Wash technology, utilizing a special plate design, has also been shown to boost the % of lymphocyte populations in your flow analysis, as a result of better debris removal and less cell loss due to centrifuge washing. Curiox also produces a smaller Laminar Wash HT2000 system, which only does washes, but provides an interesting entrance to the automated sample prep world. Lastly, there is Stratedigm, who have just gone ahead and decided that they can just automate everything. With a suite of products including an incubator, plate loader hotel, cell prep system, and high throughput plate loader system, all compatible with their brand flow cytometer. While I certainly wouldn’t throw away your pipette quite yet, this does appear to be a growing area of interest as panels become larger and more complicated
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Conclusion
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There are always new and exciting advances in flow cytometry that we in the facility want to offer to support and push forward your research. We’re always looking to bring in new products and equipment or test and evaluate new software, but it always helps to get input from our user base. If you ever see or hear about something new, or even old, technology in the flow cytometry space and would like to test it let us know! We’re happy to try and arrange demos for software or hardware when possible.
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