Aria 1 and Aria 2

BD FACSAria II

The BD FACSAria™ II Cell Sorter is a benchtop high-speed flow cytometry cell sorter with a fixed-alignment cuvette flow cell and up to 5 air-cooled lasers. It allows for high-speed multicolour sorting with digital acquisition rates of up to 20,000 events per second.

The flow laboratory is equipped with two Aria sorters. Both can sort into two- and four-way bulk sorting devices for a variety of tube sizes or can sort using Automated Cell Deposition Unit (ACDU) to multi-well plates or microscope slides. The BD FACSAria™ II Cell Sorters use FACSDIVA software for acquisition and analysis.

Configurations of the BD FACSAria™ II Cell Sorters

BRC ARIA 1 (operated by advanced and trained users only):

4 laser, 10 parameter instrument (355 nm (nUV), 405 nm (violet), 488nm (blue), 633 nm (red)), either the UV or the violet laser can be used, not both simultaneously. ACDU installed. Laser, detector and filter configuration 6 Blue/2 Red/2 Violet or nUV:

Laser	LP Mirror	BP Filter	Intended Dye
488 nm optional	735 nm	780/60 nm	PE-Cy7
	655 nm	695/40 nm	PE-Cy5.5, PerCP Cy5.5
	610 nm	616/23 nm	PE-TexasRed
	556 nm	585/42 nm	PE
	502 nm	530/30 nm	FITC
	Blank	488/10	SSC
	655 nm	675/20 nm	Pe-Cy5, PerCP
633 nm	735 nm	780/60 nm	APC-Cy7
633 nm	Blank	660/20 nm	APC
405 nm	505 nm	530/30	AmCyan
405 nm	Blank	450/50 nm	Pacific Blue
351 nm	505 nm	530/30	Indo1
351 nm	Blank	450/50 nm	DAPI/Hoechst

Note
Optional: The core laboratory owns these filters as well, but they are not routinely installed in the cytometer. If a user requires them for an experiment, please inform flow cytometry laboratory staff in advance.

Using the BRC Aria 1

The machine is user operated and available 24/7.

Only advanced flow cytometry users can access to the self service BCFACSAria II™ Cell Sorter. To become an advanced user, applicants must be approved by the senior flow cytometry manager and complete of a BD Aria operator training course.

Advanced flow cytometry users are issued with a username and password, which allows access to the sorter and the online booking calendar.

Members of the BRC's Faculty of Translational Medicine will be given priority access followed by non-members on a first-come, first-serve basis.

Flow cytometry laboratory staff reserve the right to review and/or revoke access to the self-service ARIA if a user repeatedly fails to safely operate the sorter. FCL staff also reserve the right to review and/or alter the time booked on BRC ARIA1 to allow maximum access to the sorter for all users.

Users are responsible for filling and emptying of all fluidics and for instrument setup and cleaning.

Users prepare and bring their own sheath buffer.

Book time on BRC Aria 1

BRC ARIA 2 (Flow cytometry laboratory operated):

5 laser, 20 parameter SORP instrument (355 nm (nUV), 405 nm (violet), 488nm (blue), 561nm (Yellow-green), 633 nm (red)), allows simultaneous use of all lasers. ACDU and Aerosol Management system installed. Laser/Detector and filter configuration (3 Blue/3 Red/5 Violet /2 nUV/ 5 Yellow-Green):

Laser	LP Mirror	BP Filter	Intended Dye
488 nm Optional	550 nm	575/26 nm	PE
	505 nm	530/30 nm	FITC
	Blank	4881/10 and pinhole	SSC
	505 nm	525/50	FITC
	495 nm	515/20 nm	EGFP
	525 nm	545/35 nm	YFP
	635 nm	670 LP	PerCP, PerCP-Cy5.5
561 nm Optional	735 nm	780/60 nm	PE-Cy7
	685 nm	710/50 nm	PE-Cy5.5
	635 nm	660/20 nm	PE-Cy5
	595 nm	610/20 nm	PE-TexasRed,mCherry
	570 nm	585/15 nm	PE
	750 nm	780/60 nm	PE-Cy7
	635 nm	670/30 nm	PE-Cy5
633 nm Optional	735 nm	780/60 nm	APC-Cy7
	685 nm	710/50 nm	Alexa700
	Blank	670/14	APC
	750 nm	780/60 nm	APC-Cy7
	690 nm	730/45 nm	Alexa700
	Blank	670/30 nm	APC
405 nm	755 nm	780/60 nm	QDot 800
	690 nm	710/50 nm	QDot 700
	600 nm	610/20 nm	QDot 605
	570 nm	585/42 nm	QDot 585
	505 nm	525/50 nm	AmCyan
	Blank	450/50 nm	Pacific Blue
351 nm	505 nm	530/30	Indo1
	Blank	450/50 nm	DAPI/Hoechst

Booking the BRC Aria 2

The Aria 2 is exclusively operated by flow cytometry laboratory staff.

The instrument and operators are available for sorts Monday - Friday, 10am-5pm.

All sorts are run with sterile 1x PBS as sheath buffer. Users can provide their own sheath buffer if necessary but must inform the Flow Cytometry Laboratory at least 2 days in advance.

Members of the BRC's Faculty of Translational Medicine will be given priority access followed by non-members on a first-come, first-serve basis.

The flow cytometry laboratory staff will assign sorting times and strive to avoid large blocks of unused time to maximise access. Users will be able to view the booking calendar.

To book sorting time on this instrument, send completed booking form (word) by email to BRCFlowCore@gstt.nhs.uk.

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