flowcytometer

FACS ARIA II Online Reservation: click here

Experiment setup:

·

Choose your fluorophore and check the compatibility with the Aria II filter set

·

Always have a negative control (without fluorescence stain) for optimal PMT settings

·

With multiple stained samples also have each fluorophore separately labeled to cells/beads

·

Lyse whole blood samples and wash other kind of cell samples in order to reduce background

Information:

Lasers:

·

375nm Near UV

·

488nm Blue (13mW) (Solid State)

·

633nm Red (11mW) (HeNe Air cooled)

Nozzles:

·

70mm (70 PSI typical)

·

85mm (45 PSI typical)

·

100mm (20 PSI typical)

·

130mm (10 PSI typical)

Optical filters and detectors:

Detector/Fluorophore

Bandpass filter (nm)

Location

Forward Scatter (FSC)

 

FSC Detector

PE-Cy7

780/60

Blue Octagon A

PerCP

695/40

Blue Octagon B

PE-Texas Red

616/23

Blue Octagon C

PE

585/42

Blue Octagon D

FITC

530/30

Blue Octagon E

Side Scatter (SSC)

488/10

Blue Octagon F

-

-

Blue Octagon G

-

-

Blue Octagon H

APC-Cy7

780/60

Red Trigon A

APC

660/20

Red Trigon B

-

-

Red Trigon C

Alexa Fluor 430

530/30

Near UV Trigon A

DAPI

450/40

Near UV Trigon B

-

-

Near UV Trigon C

In case your fluorophore is not amongst the list, please contact Joost or Christian to talk about the possibilities to adjust / expand the flow cytometer filters.

Sorting:

For analysis it is possible to analyze 1mL, Falcon 5mL and 15mL sample tubes.

Sorting can be done into:

·

1mL Eppendorf micro tubes (4)

·

5mL Falcon tubes (4 or 2)

·

15mL tubes (2)

·

96 well plates (1)

Flowrate can be set from 1 to 11. For sorting it is recommended not to set the flowrate higher than 6 or more than 6000 events/second. For long experiments the sample tube can be agitated with 100, 200 or 300 RPM and can be kept at temperatures of 4, 20, 37 and 42 degrees Celsius.

If cells need to be alive after sorting, larger nozzle sizes and lower sample pressure is recommended as well as buffer medium into the tubes like RPMI.

For further information or contact please mail or call Christian.