LSRII

BD LSR II
The B-D LSR II represents the pinnacle of current flow cytometry technology. Our particular LSR II is configured to provide up to 18 colors of fluorescence detection simultaneously permitting not only high order multi-parameter analysis but also flexibility in experimental design. It is also closely matched to the laser excitations provided on the iCyt Reflection sorter to permit non-sorter experimental optimization without using the more expensive sorters. The LSR II provides 7 lasers for optimizing the excitations required by a multitude of fluorochromes (see below).

HTS: Also, on the LSR II will be a high throughput sampling system. This system can sample from 96- or 384-well plates. Analysis of all 96 wells from a 96-well plate can be done in as little as 15 minutes (actual time will depend on a number of user selectable parameters). Carryover between samples is typically less than 1% (based on beads - cells may require other conditions to achieve a like or better result).

The B-D LSR II is a fully digital platform. Data from the detectors, following pre-amp processing, is sent to high resolution ADCs for linear and log display.

The LSR II will be available to be operated by Facility staff or by investigator, but will be made available initially only to those investigators who will utilize its specific capabilities. Please discuss with the Facility Director any anticipated needs you may have for the LSR II.

BD e-Learning for LSR II software: DIVA: http://www.bdbiosciences.com/services/training/elearning/index.jsp


Instrument Overview

Location: 618 Mary Ellen Jones Bldg.
Laser Excitations: 355nm, 405nm, 488nm, 532nm, 561nm, 592nm, and 640nm

  • 18 colors of fluorescence
  • FSC detection via photodiode or PMT for increased sensitivity
  • 96/384-well high throughput automated sampling

 

INSTRUMENT CONFIGURATION: Laser excitations, Possible Fluorochromes and  Long Pass - Band Pass filters:

View the 532 Tube configuration here 

Download CSV file for panel design programs

Reading filter terminology: for the LSR II, Long Pass (LP) filters reflect shorter wavelenghts to next detector in the octagon. As with all band pass filters, the two numbers designate the middle wavelength and the total band. E.g. 450/50 allows wavelengths of 425-475 through.

UV/355nm (55 mW Genesis) 2 detectors-

  1. Indo-1 (blue), Hoescht red, Fura 2, AmCyan: 505 LP - 525/50
  2. Hoescht, Indo-1 (violet), DAPI, Hoescht: 450/50


violet/406 (55 mWCUBE) -

  1. Qdot 655, BV 650: 630LP - 660/20
  2. Qdot 605, BV 605: 600LP - 610/20,
  3. Qdot 585, Pacific Orange,  BV 570 (note, exited by 561, bleeds into 650): 570LP - 585/42
  4. AmCyan, BV 500, Fixable Aqua, CFP, Alexa 430: 505LP - 525/50
  5. Pacific Blue, BV421, CFP, DAPI, Dye Cycle Violet, Cell Trace Violet, Sytox Blue, FxCycle Violet: 450/50


blue/488nm (50 mW Sapphire) -

  1. FITC, GFP, AlexaFlour488, CFSE, PKH67: 505LP - 510/20 (changed on 8/28/2012. was:525/50)
  2. PerCP-Cy5.5: 665LP - 710/50 - Note: This channel is shared with the FS PMT. They cannot be run both together.
  3. Side Scatter: 488/10


green/532nm (150 mW Compass)- PE, PE-TxRed, AlexaFluor 532, PE-Alexa610, PE-Cy5,
PE-Cy5.5, PE-Alexa700, PE-Alexa 750, PE-Cy7, PE-Alexa 647

  1. PE-Cy7: 755LP - 780/60
  2. PE-Cy5.5, PI: 685LP - 710/50
  3. PE-Cy5: 630LP - 660/20
  4. PE-Texas Red, Alexa594, Alexa 568: 600LP - 610/20
  5. PE, DSred, Pyronin-y, PKH26: 570LP - 582/15


yellow/561nm (50 mW Compass)- uses the same detection octagon as the 532 laser, but is able to excite dyes that require a longer wavelength such as dTomato or several Red and Orange Fluorescent Proteins

  1. 755LP - 780/60
  2. 685LP - 710/50
  3. mPlumb, mRaspberry: 630LP - 660/20
  4. mRuby, Alexa568: 600LP - 610/20
  5. dTomato, mStrawberry, DS Red: 570LP - 582/15


orange/594nm (140 mW Gui-UFL-200)

  1. mCherry, mRFP, max FP-Red, AlexaFluor 594, Texas Red: 600 LP - 610/20


red/639nm (40 mW CUBE)- APC, APC-AlexaFluor 700, APC-Cy7, APC-AlexaFluor 750, AlexaFluor-700, AlexaFluor-647

  1. APC-Cy7 - 755LP - 780/60
  2. Alexa Fluor 700: 685LP - 730/45
  3. APC: 675/20

 

BD LSR II