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Facility News
Happy New Year 2023 and best wishes from the FCF Staff đĽâ¨!
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How about those new resolutions that barely survive the end of the third week of January? Did you vow to always bring fresh compensation controls ? or to stop drowning cells in ethanol for the dead cell marker control ? No matter what you have decided, this year we committing to help you in any situation, so bring it on 2023 !
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In this month FACS Tips, we will be reviewing a free software that can be found online, Floreada. A good example of what the online offers can bring to the table so check it out.
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We are happy to welcome a new machine in our Biopole roster, the LSRIId. Its 4 lasers configuration can be found here and it is already available to all the BD machines users on IRIS. It is also planned for a UV laser upgrade in the coming months !
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Finally, for those who like the Cytek Aurora, there will be an online Cytek Spectral Happy Hour on Friday 10th of February 2023 from 2 to 4 pm. The details are not yet available but it will revolve around new technics and optimization of the Aurora machine. So don't forget to write it down in your calendar.
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Last month Quiz was won by Daniela Cropp! She won a Toblerone bar and the exclusive mug of the month ! We trying to bring a bit more artistic vision to those basic fluorescent proteins đ
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Each month, we will give away one of those special and unique mug designed by the FCF team so please take few minutes to answer the quiz HERE.
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FACS Tips
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Floreada
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Itâs not often that good things come free, and this is certainly true for flow cytometry. Particularly in the analysis space, FlowJo dominates with its paid subscription service along with FCS Express, both of which are available here at the FCF. There is a newly developed free alternative though! Introducing https://floreada.io/, a free to use online flow cytometry platform that is worth a try. While it may not be as robust as its paid service competitors, itâs easy to use and can be utilized in situations where FlowJo is unavailable.
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The About section on the website is quite limited, and the creators have decided not to give themselves much credit other than to describe themselves as, âExperienced low-level C programmers who are dismayed with the inefficiency of modern day commercial scientific softwareâ. Along with the About section there is a Guide section to all the steps to perform your analysis and all the tools available to you. Additionally, their support area is quite responsive as they got back to questions I had in only a couple of hours. Very nice for a free service.
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To start, simply just open the web browser and drag and drop in your files, or use the Files tab at the top to load them in. Any compensation attached to your FCS files will automatically appear underneath the sample. No worries though if there is no compensation, as it can be performed on the platform using AutoSpill. AutoSpill, which is also available in FlowJo, is an automated compensation technology (previously mentioned here in the CYTO 2022 Review Newsletter) and has been shown to create more accurate compensation values than hand gated samples, especially in weakly positive populations. All it requires is for you to load in your single stain controls and use the compensation tool on the ribbon on the left-hand side. Select your markers of interest and then make sure the AutoSpill bounds gate is in the correct position. By right clicking on one of the AutoSpill tabs beneath your sample you can display a menu. Navigate to the details section where you can find your compensation matrix, which is manually editable.
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From here you can move to your fully stained samples and gate along as you normally would. By moving gates into the pipelines area, it will be applied to all samples. Plots can be edited and adjusted, then exported for display. Itâs even possible to get overlays and batch analysis reports. All the essential features that you would expect in a paid service appear to be available. Additionally, they include some tools like clustering, cell cycle, and kinetics analysis wizards, although I havenât tested them yet. Itâs also possible to save an analysis workspace and then load it back in for future samples. Just use the File tab at the top to save GatingML Workspace, and you can then select to open that file on future analysis sessions.
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No platform is without its weaknesses though, and I have noticed some problems. Moving from my uncompensated to compensated files I lost my marker labels, and I havenât yet gotten a solution for this from the Floreada user support team. Also, if the web page refreshes, you will lose your analysis and have to start from scratch. Hard to find too many complaints on a free service.
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This can be a great solution for groups that do some flow analysis, but maybe not enough to justify paying for the FlowJo license. Or, if you were limited by a dongle and needed to do some analysis away from the lab it provides an easy, convenient alternative. While it certainly doesnât have all the features and tools of a paid subscription platform, itâs nice to use and creates a greater competition in the field. As a rising tide raises all boats, this can only make things better for users across platforms. As always, feel free to ask the FCF team if you have any questions.
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