Flow Cell Laser Intercept Movies From Development And Current Cytometer Instrumentation
FACS Aria
We do not subscribe to a maintenance agreement on our FACS Aria so service tools and jigs had to be fabricated by our Flow Cytometry Engineering department. One such device vital to re-couple the Aria flow cell is a service scope to view the actual laser incepts occurring inside the quartz cuvette and aligning proper position.
This scope could also be used on the FACS Canto.
RPCIC/HRI fabricated Aria/ Canto service scope:
CCD camera inserted in place of the above Aria service scope ocular and a video clip recorded (FACS Aria intercepts):
CMOS color camera held in place of the scope ocular and a video clip recorded
(FACS Aria intercepts):
Our last Flow Cell re-couple revealed the necessity to perform this service more often than on our non-sorting bench cytometers. After a period of 4 months, a build up of salts occurred behind the flow cell. It was not noticeable until the flow cell was uncoupled. Clean up of the mounting area and flow cell re-couple restored instrument sensitivity. The daily QC bead CVs were not overly broad but the channel values dropped 20K to 30K channels as noted before re-couple. Our theory is that persistent nozzle tip assembly leaks due to misaligned o-rings by the operator and weeping caused flow cell fluid to seep behind the assembly.
As shown above, the Flow Cell viewing scope ocular was removed and a small CCD camera is inserted. The image magnified by the objective is then focused onto the CCD element. Below is a clip of the resulting image.
Intercepts seen are 488nm (top), 408nm, 658nm, 785nm respectively.
The next movie is from the intial build of an IR (Infra-Red excitation) bench top Flow Cytometer. It is a single solid state laser (785nm 40mw, adjusted to 20mw at the Flow Cell) system with FSC, SSC and a 820LP detector (APD technology >1000nm sensitivity).
The picture shows the service scope again mounted into position to image the core stream intercept. The 820LP APD detector was removed and a light source inserted in it's place to "Back Light" the optical path. The use of a CCD camera was started with this IR project since the laser intercept is not
visible
by the human eye.
