manufacturer:       Becton Dickinson

model:                   LSR II

annotation:           Dokumente engl.

BD ^TM LSR II is the first and only air cooled four-laser benchtop flow cytometer with the ability to acquire 10 or more colors. At the heart of the BD LSR II is the new digital acquisition system, sampling signals at 10 million times a second, providing increased detection and channel resolution per pulse, thus yielding more information per cell. BD digital software provides both online and offline compensation running in a Windows®- based environment. Configure your cytometer with fixed-alignment 488-nm, 633-nm, 405-nm, and 355-nm lasers. Each laser has a spatially separated beam spot. Light is transmitted by fiber optics to an innovative Octagon detector array with user-interchangeable reflective dichroics and optically matched bandpass filters that produce increased sensitivity. Multicolor experiments have now gotten easier by running them on the BD LSR II.

Specifications*

Performance

Fluorescence Sensitivity

Estimated detection limit is <200 molecules of equivalent soluble fluorescein (using Spherotech Rainbow RCP 30-5A particles).

Fluorescence Resolution

Coefficient of variation FL2-Area of <3%, full G ₀ /G ₁ peak for propidium iodide (PI)–stained chicken erythrocyte nuclei

Fluorescence Linearity

Doublet/singlet ratio for CENs stained with PI = 1.95–2.05 detected off the 488-nm or UV laser

Forward and Side Scatter Sensitivity

Sensitivity enables the separation of fixed platelets from noise.

Forward and Side Scatter Resolution

Scatter performance is optimized for resolving lymphocytes, monocytes, and granulocytes.

Sample Acquisition Rate

>20,000 events/sec (2 x 10 ⁷ cells/mL running at 60 µL/min)

Excitation Optics

Optical Platform

Fixed optical assembly

Kinematic beam steering

Lasers

20-mW; Coherent Sapphire 488-nm, air-cooled, argon-ion laser

25-mW; 405-nm Coherent VioFlame

20-mW; 635-nm JDS Uniphase HeNe

355-nm Lightwave Solid-State laser

Beam spot size

15 ±3-µm height and no greater than 75 ±15-µm width

Emission Optics

Optical Coupling

Quartz curvette is coupled to emission lens by refractive index-matching optical gel for optimum collection efficiency.

Fluorescence Detectors and Filters

Four fixed fiber apertures

(125-µm separation)

Ten fluorescence detectors

High-performance, high dynamic range photomultiplier and red‑sensitive photomultipliers with filters

Four wavelengths detected from the 488-nm laser:

515–545 nm FITC

562–588 nm PE

675–715 nm PerCP Cy5.5 or PE-Cy5.5

750–810 nm PE-Cy7

Two wavelengths from the 635-nm laser:

650–670 nm APC

750–810 nm APC-Cy7

Two wavelengths from the 325-nm laser:

395–415 nm Indo-1 (Violet)

425–475 nm DAPI or Hoechst

Two wavelengths from the 405-nm laser:

420–460 nm Cascade Blue

505–585 nm Alexa 430

Additional bandpass filters supplied:

663–677 nm PE-Cy5

750–810 nm PE-Cy7

515–545 nm Indo-1 (Green)

Filters and mirrors are user changeable.

Signal Processing

Workstation Resolution

Five decades for peak detection: offset less than 1/10,000 of full scale

Data Acquisition Channels

Twelve acquisition channels with up to four time-displaced laser beams

Dynamic Range

18-bit data acquisition

262,144 linear channels displayed in a 5-decade logarithmic display

Fluorescence Compensation Networks

Inter- and intra-beam software- programmable resolution of 0.01%

Pulse Processing

Width and Area measurements available for all fluorescence parameters

Ratio measurements for intra-laser parameters (useful for calcium measurements)

Time

Time available correlated to any parameter

Channel Trigger

Triggering on any laser signal

Ability to create logical AND/OR of channel triggers

Ratio

Ratio measurements between any two parameters

Fluidics

General Operation

Front key panel provides three modes: RUN, STNDBY, and PRIME;

automatic standby mode conserves sheath fluid by stopping sheath flow when no sample tube is installed.

Sample Flow Rates

Three selectable flow rates

Regulated pressure difference between sheath and sample

Quartz Curvette

Internal cross-section is rectangular 430 x 180 µm; external surfaces are anti-reflection coated for maximum transmission of laser light.

Data Management System

Workstation

HP X4000 workstation or equivalent

Central Processing Unit (CPU)

Dual Intel Xeon processors

Memory

2 GB RD RAM

Data Storage

Nine-and 36-GB internal hard drive, internal drive with 250-MB removable disk storage space capacity, 3.5-inch floppy drive, CD-RW drive, tape drive

Networking

Ethernet

Monitor

Two 18-inch LCD monitors

Printer

Networkable color-inkjet printer

Data File Structure

Flow Cytometry Standard (FSC) 3.0 or 2.0