- LSE Center Publications
- Microscopy & imaging
- Super Resolution Microscope: Lattice SIM/STED/STORM
- Zeiss- Light Sheet Z7
- Nikon- Spinning Disk Confocal
- LSM 880- Upright confocal with MP laser
- LSM 710 – Inverted confocal
- LSM 700 – Inverted confocal
- LSM 980- Inverted confocal with Airyscan2
- Cytation™ 5 – BioTek
- Leica DMI8 Inverted Fluorescent Microscope
- Olympus Fluorescent Binocular Microscope
- Image analysis & processing
- Flow Cytometry
- Virus room – Emerson Building
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 thresholds 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
|
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,
|
|
| E | 488 / 10 (Blue) | SSC | |
| FSC | |||
|
633 nm Red
|
A | 780 / 60 (Infra Red) | APC-Cy7, APC-H7 |
| B | 660 / 20 (Red) | APC, Alexa Flour 647 | |
|
405 nm Violet
|
A | 695 / 40 (Infra Red) | BV650, BV711 |
| B | 450 / 50 (Blue) |
Pacific Blue, BV421 |
|
|
355 nm UV
|
A | 530 / 30 (Green) | Indo 1 (Blue, Calcium Flux) |
| B | 450 / 50 (Blue) | DAPI , Hoechst, Indo 1 (Violet) |
-
High Throughput System (HTS) – enables analysis directly from 96 multi-well plate. The workstation is managed by FACS-Diva 6.1 software.
Additional available filters:
| Mirror | 450LP | 5005LP | 530LP | 635LP | 685LP | 735LP | ||
| Band pass | 405/20 | 525/50 | 530/30 | 585/42 | 610/20 | 670/14 | 675/20 | 695/40 |
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
Related Documents