CytoFLEX Cell Counter / Flow Cytometer to be installed installed Tuesday 30th October 2018!
We will be installing the recently acquired CytoFLEX next week and will soon update on access guide to using the new instrument. The CytoFLEX provides volumetric acquisition providing cell counting to users.
Training for key SuperUsers will occur on Friday 2nd November with an access workflow finalised soon after for users to start using it.
Left: Cytoflex Right: Ian & Mathi post Cytoflex install
Christmas Closedown
The Westmead Institute will not be closing over this year’s break; please note there will be less support services and reduced building access hours from 8am-8pm each day.
Please note that Westmead Cytometry will not be serviced from Monday 24th December 2018 (Christmas Eve) to Tuesday 1st January 2018. There will no QC run on the flow cytometer analysers if there are bookings during this period. Please consider this and ensure a QC has been run before running your samples during this period if you need to use the instrument. The sorters will not be operational on Friday the 21st of December as they will be shut down for the Christmas Closedown. The sorters will also not be open for bookings on Wednesday the 3rd of January as they will be brought back online on this day. The first available day for a sort for researchers will be the 3rd of January 2018.
There will be reduced facility support in place during this period and it is requested that only essential research work is undertaken.
'WRHFlow' to transition to 'Westmead Cytometry' in 2019
We will migrate to using westmead.cytometry@sydney.edu.au over the next few months but will continue to monitor both the new email address as well as the existing wrh.flow@sydney.edu.au email address.
8 Peak Beads – Now available to users
8 peak beads are now available for users to acquire as a means of QC on the WRHFlow cytometers. Users can find the 8 peak beads alongside the QC(CST) beads in the flow cytometry fridge in the shared lab. Acquiring 8 peak beads before and after your acquisition provides information on whether the instrument has changed from the start to the end of your run. They can also be used to set/monitor PMT signals between acquisitions performed on different days in synergy with application setting for those users who are using application users. Please ensure you record the bead lot number!
Instrument Baselines – November 2018
From 2017 to the current, we have maintained the same target MFIs on the 2 CantoIIs and the Fortessa via an almost daily quality control (note: LSRII baseline had been updated earlier in 2018). This has allowed users to utilise application settings for longitudinal studies. The below notice is for users currently using application settings on the instruments.
As the current QC beads in use will finish in the next couple of months we are required to establish new target MFIs (also known as baselines) for the new bead lot that has arrived. This is also necessary when major changes have occurred on the instrument. Major notable changes that have/will occur include laser, computer, and electronic component replacements on different instruments.
To those users not using application settings there will be no further disruption.
To those users using application settings and who wish to continue with existing target MFIs, know that you will need to save new application settings moving forward as we will record new baselines progressively in November 2018 as per the below schedule. This needs to be done once only.
The recommended method that would work for users who have not recorded beads in prior experiments, would be to switch to the existing baseline (the one that existing application settings were saved on), run a CS&T using the previous bead lot, make a new experiment, apply existing application settings, copy the voltages, switch back to the new baseline, run a CS&T using the new bead lot, make a new experiment, copy the voltages over to the new experiment, and save new application settings. If any help is required please contact WRHflow well in advance as a mistake would propagate to other users. The method is outlined below.
*Note for CantoII & LSRII you will need to change to an existing configuration. Please contact staff if help is needed.
1) Run CS&T > Cytometer > CST > Performance check > using bead lot #70466 (old)
2) Exit CS&T, make a new experiment, apply your existing application settings.
3) Record your voltages!
4) Run CS&T > Cytometer > CST > Performance check > using bead lot #80998 (new)
5) Exit CS&T, make a new experiment, manually update your voltages.
6) Save new application settings with a distinctive name.
7) Your application settings will now be linked to the new bead lot number which we expect to last for ~ 1 year.
It is important to ensure the above process is completed once started – to ensure the most recent CS&T report is from the #80998 bead lot, as the instrument uses the last CS&T data acquired.
Unfortunately the above method will not be applicable to the LSRII as we are planning a computer/software change. LSRII users are recommended to run and record 8 peak bead data (and record the bead lot) referencing peak locations in order to carry over any detector voltages if necessary.
New Baseline Implementation Schedule
...
Instrument
...
Date Scheduled
...
Bead Lot
...
CantoII_ICPMR
...
5/11/2018
...
#70466 > #80998
...
CantoII_WIMR
...
12/11/2018
...
#70466 > #80998
...
LSRII
...
31/10/2018
...
#70466 > #80998
...
Fortessa
...
19/11/2018
...
#70466 > #80998
...
Symphony
...
17/9/2018
...
#70466 > #80998
...
Did you say colourful? BD Symphony is outputting high quality data.
The Symphony is ready to use! The instrument has been used by a number of researchers with positive feedback. The current configuration allows for at least 34 (yes 34) different fluorescent dyes to be detected in addition to 3 scatter parameters. Be sure to check out the current configuration listing the non-exhaustive plethora of dye choices.
Why would you want to use it? Well, if you are having trouble choosing fluorochromes for your panel, the increased options certainly may help, or if you are finding it difficult to find time on the Fortessa/LSRII, all currently utilised flurochromes can now be detected on the Symphony ensuring an event-free (*no pun intended*) transition. The additional scatter parameter can help also increase dynamic range allowing both small and large particles to be detected simultaneously.
Training takes ~30minutes if you already cytometer access and requests are accepted through PPMS.
Fancy Using A Supercomputer? I did (and loved it).
Argus is the University of Sydney’s “Virtual Remote Desktop” project designed to bring remote, on-demand, interactive, graphically intensive compute environments to researchers.
Westmead Cytometry & Westmead Imaging now provide access to 3 of these systems for you to access for those compute intensive applications. These remote desktops are fast. Each system has 16x XEON core CPUs and an impressive 128GB RAM alongside 8GB GDDR5 NVIDIA GPUs with network throughputs of 200+ GBPS. NBN is 1GBPS for comparison.
The Argus team (special thanks to Ashok Pachipala @ USYD) have installed all of the right applications including R, RStudio, Huygens, ImageJ, CellProfiler, FlowJo(T&Cs apply), Office, AutoCAD and others making it an excellent desktop to log into, setup something that will take a while (tSNE, Image Deconvolution, etc), and let it do its thing.
Access is easy, all that is required is a Unikey and a return email acknowledging some basic guidelines. Booking is via PPMS and shhhh… they aren’t charged. These desktops have proven so capable that they even have altered mindsets of purchasing new analysis PCs.
Have Something You Need *3D* Printed?
A reminder that we have a 3D printer that can be accessed by all researchers. It has proven very capable with a number of useful prints having been produced to date. Slide holders, electron microscope parts, centrifuge parts, kidney transplant mounts, and a build your own cytometer. If you have a requirement for a custom designed novel scientific part that will assist your research, let us know!
More information can be found here
An Automatic Plate Washer!
It has been brought to my attention that many researchers are in the dark that we have an automatic 96 well plate washer available in the flow labs. If you could use this for plate based assays, such as ELISA’s, or protein quantification assays, then it is very easy to use. If you would like to be trained to use it training requests are through PPMS.
For a complete list of instruments available at Westmead Cytometry click here
Cell Counter - Now Available – Give It A Try.
Still counting PBMCs or splenocytes manually?
The cell counter can also. 50uL and ~<1min/sample.
The newly installed CytoFLEX (2 laser, 4 parameter) flow cytometer is available to use @ Westmead Cytometry. The instrument can be used as an accurate cell counter providing researchers with an accurate, quick, accessible cell counter to utilise in your research. The instrument doesn’t require booking and training is via key superusers located across the WRH. In addition to cell counting, parameters such as GFP expression, viability and scatter profiles can be collected. The instruments accepts Eppendorf tubes and 5mL FACS tubes, requiring 50uL and ~40seconds per sample. Facility provided DAPI and Syto Dyes are available also.
Key SuperUsers who can be contacted for training/further information include:
Suat Dervish
Edwin Lau
Ya Wang
Joey Lai
Cindy Zhu
Stephen Schibeci
Daniel Hu
Nicole Fewings
Cyto2019 – Vancouver - Abstract Submission Is Now Open
Cyto is unquestionable the most prominent cytometry research conference internationally. Cyto2019 will be held in Vancouver, June 22 – June 26 – 2019.
Submit an abstract before February 11th for your opportunity to present your data to an audience of cytometry/immunology/many discipline experts (an eclectic bunch). Just as important is what can be gained from the conference, i.e. the latest in data analysis workflows /algorithms, latest reagent/assay information, and sneak peaks of novel disruptive technologies entering the cytometry space. Travel awards are available for eligible applicants.
Scientific Platforms Data Clean Up Reminder
Some of you are probably using the Scientific platforms drive to store you exported flow data temporarily. As the drive is getting full, we will have to clean up the drive according to our data policy: All data >14 days old will be deleted.
On 15/2, IT will be deleting data files on Scientific Platforms that are more than 14 days old automatically. Please save all wanted files onto your local storage.
If you have any concerns please contact us.
Help Out A User 😊
If you can assist with Cindy’s request please contact her via - c.kok@sydney.edu.au
“I’m looking for a CD44 antibody that reacts to either human or rat. Could you please help me to ask around to see if anyone could spare an aliquot of 1-2 uL?”
If you can assist with Jason’s request please contact him via - jher8412@uni.sydney.edu.au
“I was hoping you would be able to help me search for a Daudi cell line. My group is interested in investigating NK cell ADCC of cell lines after stimulation with viral antigens. For this we would like to acquire some Daudi cells and if possible we were hoping there may be a group within the Westmead community that may have these cells. Could you please circulate this request through the Westmead community.”