Introduction to the LSR II and FACSDiva 6.0 Software.

Introduction to the LSR II and FACSDiva 6.0 Software

Four-laser, 10-color benchtop flow cytometer

High Throughput Sampler

Laser: 488nm, 355nm, 405 nm, 633 nm

left cover right coverside door

control panel SIP power switch

fluidicsinterconnects

Control Panel

SAMPLEFINE ADJ

LO MED HI

RUN STNDBY PRIME

sample flow ratefluid control buttons

fluid control buttons

Sample flow rates: approximately 12, 35, and 60 μL/min ( LO, MED, and HI settings )

355 nm (UV) : Indo-1, DAPI, Alexa Fluor 350405 nm (Violet) : CFP, Alexa Fluor 430, Pacific Blue, Marina Blue, Alexa Fluor 405488 nm (Blue) : FITC, PE, PerCP, Pe-Cy5 (aka CyChrome), PI, PerCp-Cy5.5, PE-Cy7, GFP, PE-Texas Red633 nm (Red): APC, APC-Cy7, Alexa Fluor 647, Alexa 680

bandpass filters

longpass dichroic mirrors

Octagon Detector Arrays

488-nm blue laser

PE-Cy5

633-nm red laser

660/20

APC

735 LP

780/60

APC-Cy7

670/14

PerCP-Cy5.5

635 LP

FITC530/30505 LP

PE-Cy7780/60

735 LP

PE

575/26

SS

C488/10

PerCP

GFP

PE-Cy5

695/40BP685LP

PI

355-nm UV laser

450/50

505 LP

530/30

DAPI

Indo-1 (Blue)

405-nm violet laser

440/40

Pacific Blue

505 LP

525/50

AmCyan

Alexa405

Alexa350

CascadeB

CascadeY

To Login the computer

Login Name: flow user Password: fccf

shortcut icon

To start the software, double click the shortcut icon on the desktop

Login to FACSDiva Software

FACSDiva workspace

Preparing Sheath and Waste Containers

Fluidics CleaningEach time you finish using the cytometer, clean the sample injection tube and the area between the injection tube and the outer sleeve. This prevents the sample injection tube from becoming clogged and removes dyes that can remain in the tubing. CLEANING INSTRUCTIONS ARE POSTED ON THE LSRII

LSRII Location:

BioSquare III, 670 Albany St. 5th Floor

(Card access is required to enter the room)

FCCF web address:

http://www.bu.edu/cores/flow-cytometry

FCCF email:

[email protected]

Sorting facility phone number:

617-414-5225