Instrument Details (Glasgow Celesta)

3.1. Instrument Manufacturer

BD Biosciences

1030 Eskdale Road, Winnersh Triangle, Wokingham, Berkshire, RG41 5TS

Website: http://www.bd.com/

3.2. Instrument Model

BD CelestaTM Flow Cytometer, Serial number H66034500019

3.3. Instrument Configuration and Settings

3.3.1. Flow Cell and Fluidics: The instrument has not been altered; fixed-alignment cuvette flow cell. 3.3.2. Light Sources: The instrument has not been altered; three-laser base configuration:

• 488-nm Bioray solid state laser; 20 mW

• 561-nm Coherent Obis solid state laser; 50mW

• 405-nm Bioray solid state laser; 50 mW

3.3.3. Excitation Optics Configuration: The instrument has not been altered.

3.3.4. Optical Filters: See figure below.

3.3.5. Optical Detectors: The instrument has not been altered.Photomultiplier tubes, Matsusada Prescision Inc. Osaka Japan. See detector voltages below (User to complete)

3.3.6. Optical Paths: The instrument has not been altered. Detector arrays consist of a BD octagons (488 nm, 561 and 405 nm laser lines). PMTs are arranged as follows: Laser PMT Filter LP Mirror Fluorochrome Blue Octagon 488nm A 695/40 670LP PerCP-Cy5.5 >20mW C 530/30 502LP BB515, FITC, AF488, GFP, YFP D 488/10 None SSC Yellow/ Green Octagon 561nm A 780/60 750LP PE-Cy7 >50mW B 670/30 635LP PE-Cy5, PI, 7AAD C 610/20 600LP PE-CF594, PE- Dazzle, mCherry D 586/15 None PE, DSRed, TdTomato Violet Octagon 405nm A 780/60 750LP BV785 >50mW B 710/50 690LP BV711 C 670/30 655LP BV650 D 610/20 595LP BV605 E 525/50 502LP BV510, V500, L/D Aqua, AmCyan F 450/50 None BV421, V450, PacBlue, eCFP, tag2 BFP

LASER PMT Filter LP Mirror Fluorochrome Blue Octagon A 695/40 655LP PerCP-Cy5.5 488nm C 530/30 502LP FITC, GFP, CFSE, AF488 >20mW SSC D 488/10 None Yellow/Green Octagon 561nm A 780/60 735LP PE-Cy7 >50mW B 710/50 685LP PE-Cy5.5 C 660/20 635LP PE-Cy5, PI, 7AAD D 610/20 600LP PE-CF594, mCherry, PE- Dazzle, 7AAD E 585/15 None PE, DSRed, AF546

Red Trigon 640nm A 780/60 755LP APC-Cy7 >40mW B 730/45 690LP AF700 C 670/14 None APC Violet Octagon 405nm A 780/60 750LP BV785 >50mW B 710/50 685LP BV711 C 660/20 630LP BV650 D 605/20 595LP Qdot 605 E 525/50 502LP AmCyan, V500, Live dead aqua F 450/40 None DAPI, V450, Pac Blue, eCFP