BD - LSR Benchtop Flow Cytometer

 

The JCSMR Flow Cytometry Unit "proudly presents" the Becton Dickinson LSR Benchtop Flow Cytometer. The LSR is a modified 6 colour, 4 laser Flow Cytometer.

Following are some of the features and specifications of the LSR.

Lasers: The LSR is equipped with four lasers. The primary laser being an argon-ion 488nm with 20mW power output. The second laser is a Helium-cadmium 325nm (UV) with 8mW power output, the third is a Helium-Neon 633nm (red) 17mW power output laser and the fourth laser is a 594nm (yellow) Helium-Neon.

Fluorochromes and Fluorescent proteins that are suitable for the LSR.

Laser Fluorochromes
488nm argon-ion FITC, PE, PE-Cy5(Tricolor/Cychrome), PE-Texas Red, Propidium Iodide, Acridine Orange, Ethidium Bromide, Rhodamine, 7-AAD, PerCP, GFP and YFP
325nm Helium-Cadmium UV Alexa350(Being tested), Indo-1, Hoechst33342, Dapi
633nm Helium-Neon APC, APC-Cy7
594nm Helium-Neon Alexa 594, Texas Red

 

This table shows only the more common fluorochromes, for more information about fluorochromes go to http://pingu.salk.edu/fcm/fluo.html or http://facs.scripps.edu/spectra or www.probes.com

Detectors and Filters: There are 6 fluorescence detectors and 2 scatter detectors in the LSR.

 

Detector
Steering Optics (mirrors)
Filters
  
SSC
488/25 BP
FCS
488/10BP
FL1
555 DRLP
530/28BP
FL2
575/26BP
FL3
620SP and 670LP
670LP
FL6 (SSC-W)
670/40BP*
FL4

510LP and 470LP
510/20BP
FL5
424/44BP

* Non standard Filter

Other Filters available are:

  • 400/40nm for indo-1 (bound Ca++)
  • 380nmLP UV eg. Dapi and Hoechst33342
  • 610/20nm PE-Texas Red
  • 682/13nm can be used for PerCP
  • 660/13nm can be used for APC
  • 780/60nm can be used for APC-Cy7

 

For a look at the default Filter layout and filter combinations for different fluorochromes, click here !! (under construction!!)

Filter glossary

SP ==> Shortpass Filters transmit wavelength that are shorter than the specified wavelength.

LP ==> Longpass Filters transmit wavelength that are longer than the specified wavelength.

BP ==> Bandpass filters transmit a well-defined band of light, and reject all other unwanted light.

DF ==> Discriminating Filters are a type of bandpass filter.

DRLP ==> Dichroic Longpass Filters (mirrors)transmit longer wavelenth to one detector while reflecting shorter wavelength to a different detector

DCSP ==> Dichroic Shortpass Filters (mirrors) transmit shorter wavelength of light to one detector while reflecting longer wavelength to a different detector.

Flow rates:

HI
 30ul/min - 120ul/min (five turns of sample fine adjust button = midpoint of 60ul/min)
MED 17.5ul/min - 70ul/min (five turns of sample fine adjust button = midpoint of 35ul/min)
LOW 6ul/min - 24ul/min (five turns of sample fine adjust button = midpoint of 12ul/min)

 

Click here for a closer look!

 

Data acquisition and analysis:

The data acquisition software is CellQuest pro. For more information about new features of CellQuest pro compared to the old CellQuest, have a look at the "What's new movie" . (Requires Quicktime)

Here are some datafiles generated on the LSR. (Note: Except for the indo-1 Calcium files, alignment Beads were used.)

Examples of Ca++ flux with indo-1 staining of mouse lymphocytes, with time as a parameter on the x-axis and the FL5 (400/40nm) vs FL4 (510/20nm) ratio on the y-axis.

 

A cooling/heating system with a temperature range from 4 degrees - 120 degrees is connected to the system via a water jacketed tube to provide sample temperature control.

GLOSSARY

Links to more information about the LSR

 

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Created by Sabine Grüninger , last updated 1-11-01