BD LSR-II Analyzer

The BD LSR-II is a flow cytometer equipped with 4 lasers. The cytometer allows detection of 12 parameters simultaneously (10 colors plus forward scatter and side scatter).

The LSR-II offers high acquisition speeds, full inter- and intra-laser compensation matrix, and multiple thresholding on all lasers. Digital pulse sampling allows the measurement of Height, Area and Width from all parameters simultaneously.

 

Features

  • Optical System: Patented BD OctagonTM and TrigonTM technology. A quartz cuvette is coupled to the emission lens by refractive index-matching optical gel for optimum collection efficiency.
  • laser excitation and filters (see LSR-II configuration in the related links):

 

 

Laser line/color
(Excitation)
Detector Emission Filter (nm) Example of available markers

488 nm / Blue

  Coherent Sapphire
solid-state laser,

  22 mW

 A  780 / 60 (Infra-Red)  PE-Cy7
 B   695 / 40 (Red-FarRed)

PE-Cy5, PerCP, PerCP-Cy5-5,

  7-AAD

 C  575 / 26 (Orange-Red)  PE, PI, PKH26
 D   530 / 30 (Green)

Alexa Four 488, FITC, GFP,

  CFSE, Sytox Green, YFP,

  BOBIPY, Oregon Green,

  Pico Green

  E (SSC)  488 / 10  SSC
 FSC  FSC

633 nm / Red

  JDS Uniphase Helium
Neon laser, 20 m

 A   780 / 60 (Infra Red)   APC-Cy7, APC-H7
 B  660 / 20 (Red)  APC, Alexa Flour 647

405 nm / Violet

  Coherent VioFlame
solid-state laser,

  25 mW

 A   525 / 50 (Green)  AmCyan, Cascade Yellow
 B   450 / 50 (Blue)

Alexa Flour 405, Pacific Blue,

  Cascade Blue

355 nm / UV

  Lightwave solid state
UV laser with 20 mW
at 355 nm

 A  530 / 30 (Green)   Indo 1 (Blue, Calcium Flux)
 450 / 50 (Blue)  DAPI , Hoechst, Indo 1 (Violet)

 

ndividual filters and PMTs, allowing for 11 color analysis simultaneously (see configuration for more details)

 

  • High Throughput System (HTS) –  enables analysis directly from 96 multi-well plate

The workstation is managed by FACS-Diva 6.1 software.

 

Specifications and performance

Acquisition Rate: >20000 events per second

Fluorescence Sensitivity: Detection limit is <200 molecules of equivalent soluble fluorochrome (MESF).

Fluorescence Resolution: Coefficient of variation (CV) of FL-Area is <3%, determined by full G0/G1 peak for propidium iodide (PI)-stained chicken erythrocyte nuclei (CEN Quality Control Kit).

 

Fluorescence Linearity: Doublet/singlet ratio for CENs stained with PI or Hoechst is 1.95-2.05, detected with 488nm or UV laser.

 

Forward and Side Scatter Sensitivity: Sensitivity enables the separation of fixed platelets from noise.

 

Signal Processing

  • Digital electronics with 10 MHz sampling frequency
  • 18-bit data acquisition for area signals
  • Inter- and intra-beam compensation on-line and post-acquisition off-line
  • Height, Width, and Area measurement available for all parameters
  • Time parameter available for kinetic studies (Calcium)
  • Ratio measurements between any two parameters from any laser

 

Software

  • BD FACSDiVaTM Software Version 6.1
  • All data are saved linearly
  • Complex hierarchical gating for multicolor applications
  • Automatic compensation of all parameters after acquisition
  • Complete set of statistics for all analysis

 

Applications

Analysis of cells, yeast, bacteria and beads

  • Multi-color analysis
  • HTS system
  • Calcium Flux
  • DNA analysis

 

Releted Documents

BD LSR-II User Guide

BD LSR-II Configuration

Selecting Colors For Multi-color Flow Cytometry

BD FACSDiva Software 6.0 Manual

 

Releted Links

BD-LSR-II