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- #How to compensate in flowjo 10 in window for mac
- #How to compensate in flowjo 10 in window update
- #How to compensate in flowjo 10 in window software
- #How to compensate in flowjo 10 in window license
#How to compensate in flowjo 10 in window for mac
WRHFlow recommends calculating the spillover spreading matrix using a recent version of FlowJo - v10 (FlowJo v9 for MAC also has the functionality).
#How to compensate in flowjo 10 in window software
20180621 - What software can I use to calculate a spillover spreading matrix? A universal negative will only use events that fall under the P1 gate. It then also requires a P2 population for positive and (if not using a universal negative) a P3 population for negative events for each fluorochrome. The software looks for a P1 gate on a bivariate FSC and SSC plot. 20180724 - Why can I not calculate compensation on DIVA? If you haven't, follow this link ( ), complete the quiz, and email access. If you have accumulated 10hrs or more of experience (used time) on the instrument we can instantly grant you autonomous access.
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20180913 - Can I get autonomous (after hours access - still subject to building access - on the flow cytometers)? Trying AnnexinV on colours other than the commonly used FITC (APC, BV421), as well as using DAPI instead of PI (not if using AnnexinV BV421) can also provide better resolved populations.
#How to compensate in flowjo 10 in window update
The online versions are update however in the interim.Ĭytometer Configurations 20181009 - What comp beads can I use for the Annexin/PI kit for apoptosis detection.įor PI/AnnexinV kit, there are no beads that bind PI, so actually using cells would provide adequate compensation for both AnnexinV and PI. We are currently renewing baselines and will have the printouts up soon by the instruments. All we need is the hardware address of your new computer.Ģ0181105 - Where did the cytometer configurations go?
#How to compensate in flowjo 10 in window license
20190129 - Can I transfer my FlowJo license to a new computer? NA/LE format) but commonly for research cell sorting this isn't common. You can purchase endotoxin tested and azide free antibodies (eg. The current idea is that the concentration (once cells are stained) is low, followed by all the washing to remove unbound antibody that their is minimal azide that remains with the cells. This is an excellent quest ion and the answer is dependent completely on the researchers requirements.Ĭlose to all current Westmead Cytometry researchers who are sorting their cells for downstream functional assays do not specifically stain with azide free antibodies. Most pre-conjugated antibodies contain sodium azide to minimise bacterial contamination. 20190122 - Do we need sodium azide free ABs for cell sorting? If the spillover is high then yes FMOs would be useful. There can be a use for these in determining gating positions, depending on the amount of fluorochrome spilling over into other channels.
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The 2 colour samples in this scenario could be called FMO controls (fluorescence minus one controls). I plan to have a no stain, and single stain controls. Jun-Aug 20190304 - We are planning a three colour FACS Canto experiment. The FlowJo site licence billing cycle starts on the 1 st of September and the quarters are as follows.
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20190326 - Do you know when the next billing cycle is for the FlowJo site license option at Westmead Cytometry? FAM (essentially FITC) is excited well by the 488nm laser and again emits with a slightly longer wavelength that can be detected with the FITC detector on the CantoII. I’m wondering if it might be possible to do this on the CantoII? Or do I have to split my colours up into two experiments?ĪF647 is excited well by the red ~633nm laser and emits at a slightly longer wavelength. I have LDS-751 and EdU-click conjugation with AF647 and PNA-probe conjugated to FAM. 20190517 - I am planning to do Flow-FISH using 3-colour system.