Flow Cytometry and Cell Sorting
Instrumentation: BD LSR Fortessa
BD LSR Fortessa
Location: The Ottawa Hospital Sprott Centre for Stem Cell research, 5th floor Critical Care Wing, room W5215, 501 Smyth Road
The BD LSRFortessaTM cell analyzer offers the ultimate in choice for flow cytometry, providing power, performance, and consistency. Designed to be affordable and expandable, the BD LSRFortessa has the flexibility to support the expanding needs of multicolor flow cytometry assays.
Optics specifications:
The BD LSR Fortessa is equipped with 5 lasers (355nm, 405nm, 488nm, 561nm, 633nm). Arrange in octagon- and trigon-shaped optical pathways, their novel design efficiently maximizes signal detection and increases sensitivity and resolution. This allows researchers to identify cells, especially dim and rare cell populations, optimizing multicolor assays and panel design for superior results.
This configuration allows us to detect up to 18 different fluorescent parameters: 3 parameters from 355nm laser, 5 parameters from 405nm laser, 2 parameters from 488nm laser, 5 parameters from 561nm laser and 3 parameters from 633nm laser.
| LASER |
FILTER |
FLUOROCHROME |
| UV (355nm) |
379/28 |
BUV395 |
| 670/25 |
BUV661 |
| 740/35 |
BUV737 |
| Violet (405nm) |
450/50 |
BV421, V450, Pacific Blue |
| 525/50 |
BV510, V500, Am Cyan |
| 660/20 |
BV650 |
| 710/50 |
BV711 |
| 780/60 |
BV786 |
| Blue (488nm) |
530/30 |
FITC, GFP, YFP, Alexa Fluor 488, BB 515, Venus |
| 710/50 |
PerCP-Cy5.5 |
| Yellow-Green (561nm) |
586/15 |
PE, Ds Red, TdTomato, Alexa Fluor 546 |
| 610/20 |
PE-Texas Red, m Cherry, PE-Alexa 610, PE-CF594, PI |
| 670/30 |
PE-Cy5, 7-AAD, PE-Alexa 647 |
| 710/50 |
PE-Cy5.5 |
| 780/60 |
PE-Cy7, PE-Alexa 750 |
| Red (633nm) |
670/14 |
APC, Alexa Fluor 647 |
| 730/45 |
Alexa Fluor 700 |
| 780/60 |
APC-Cy7, Zombie NIR |
Fluidics:
The fluidics system is pressure driven. Hydrodynamic focusing forces sample cells through the cuvette flow cell, where they are interrogated. The flow cell is in fixed alignment with the laser and gel-coupled to the collection optics. This design ensures that the laser is precisely focused on the sample stream and the maximum amount of emitted light can be collected for added sensitivity in multicolor applications. Fixed alignment also minimizes startup time, improves experiment-to-experiment reproducibility, and enables automated daily quality control.
Fluidic sensors maintain constant pressure, while a fluidics monitoring system warns when sheath fluid is low, empty, or when the waste container is full.
BDTM High Throughput Sampler Option
To improve experimental workflow, the optional BDTM High Throughput Sampler (HTS) provides rapid, fully automated sample acquisition from 96- and 384-well microtiter plates. In high-throughput mode, the HTS option can speed through a 96-well plate in fewer than 15 minutes with less than 0.5% sample carryover from one well to the next. Low carryover is essential in research applications to ensure sample purity and data integrity.
Fast acquisition speed is achieved by synchronizing two high-precision pumps for sample mixing, sample injection, and probe washing. Standard throughput mode can be selected for acquisition of larger sample volumes.
