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The LSRII is a Special Order Research Product (SORP) digital cell analyser equipped with a Violet (405nm, 100mW), Blue (adjustable 488nm, 80mW; maximum output 100mW), Green (532nm, 150mW) and Red (642nm, 40mW) laser, capable of measuring up to 18 fluorescent parameters in addition to FSC and SSC. N.B. The warm up time for the instrument’s lasers = 30min. If you are the first user of the day, but are not based in the JR, please call ahead and we can switch on the instrument for you.

The LSRII has the optional High-Throughput Sampler (HTS) module, offering automated sample acquisition from 96-well plates; the standard mode is capable of processing a 96-well plate in under 30min (< 0.5% carryover).

This instrument was purchased by the NIHR Oxford Biomedical Research Centre (NIHR OBRC).

 Special Order Research Product (SORP)

The fluorochromes listed in the table below are examples of reagents which can be measured in each detector, but does not represent an exhaustive list. Please ask if you are unsure whether a particular reagent can be detected or not.

Violet Blue Green Red
A.       BV785/786 A.       PerCP, PerCP-Cy5.5 A.       PE-Cy7 A.       APC-Cy7, APC-AF750, APC-H7
B.       BV711 B.       FITC, AF488, BB515 B.       PE-Cy5.5 B.       AF700
C.       BV650 C.       PE-Cy5 C.       APC
D.       BV605 D.       PE-CF594

E.        QD570

E.        PE
F.        QD565*
G.       BV510
H.       BV421
* The LSRII can be set up to measure QD545 in this detector using a different software configuration (requires filter swap).
 
If you wish to measure any fluorescent parameters which are not listed above, please contact the Facility Manager  as we may have additional optical filters to fit your requirements. A more detailed instrument configuration including the dichroic mirrors and bandpass filters can be found below.

General information

  • Sample acquisition: >15,000 events/sec (2x10cells/ml running at 60μl/min)
  • The ‘Sample fine adjustment knob’ allows you to adjust the sample flow rate to intermediate levels. When the knob is at its mid-point: LO = 12µl/min, MED = 35μl/min, HI = 60μl/min. Five full revolutions from the mid-point provide 0.5-2X the mid-point rate e.g. sample flow rates of 6-24µl/min are possible when the LO button is selected.
  • Allow at least 5 minutes for the pressure to stabilise after priming the instrument – this is crucial prior to running the CS&T as not doing so could result in erroneously calculated laser delays and suboptimal data.
  • Unless the HTS is attached, under no circumstance may the outer needle be removed when running samples.

LSRII SORP configuration:

Laser

PMT Dichroic mirror Bandpass filter Range, nm
Violet

A

B

C

D

E

F

G

H

750LP

685LP

635LP

600LP

570LP

557LP

505LP

-

785/60

705/40

660/20

610/20

585/42

560/40

525/30

450/50

750-815

685-725

650-670

600-620

570-600

557-570

510-540

425-475

Blue

A

B

SSC

670LP

505LP

-

685/35

515/20

488/10

670-703

505-525

483-493

Green

A

B

C

D

E

740LP

685LP

635LP

600LP

-

780/60

710/50

660/20

610/20

585/42

750-810

685-735

650-670

600-620

564-600

Red

A

B

C

740LP

685LP

-

780/60

710/50

670/14

750-810

685-735

663-677

(Image used courtesy of BD Biosciences)