Video link - http://tbit.ly/eVhMW0
Another big day began at an early 4:37am with an excited core facility manager eager to return to the lab and continue building the BD Influx™. 7:21am I arrived at the lab and began clearing space for the day ahead, Mark and Brent arrived shortly after and we launched into it again.
In the wise words of an old friend 'Fluidics First' is exactly how today began by working on getting the jet up and running straight away. Mark worked on the compressor and Brent began on the optics. Once the compressor was in place, the power supply for the UV laser was hooked up with large power cables thick enough to swing from. Labels were printed for the 532 and 561 lasers as the sheath tanks were filled up and the fluidics was primed. The all-too-familiar hum of a mildly deafening compressor began vibrating away which was soon drowned out by the jibbering vacuum pump, it wasn't long before we began making alternative arrangements to run both vacuum and air off the house which makes a significant difference to noise reduction (note to potential customers; run house air and more importantly house vacuum). As we stepped down the pressure from the wall to the tip, each point being 20 psi less than its predecessor the sheath pressure valves soon read 90 psi and the vacuum guage read a steady 5. We began tweaking the pressure setttings on the instrument at the sample differential points and discussed nozzle tip options (70um/86um/100um/140um/200um).
In Influx fashion; simple and seamless, we boot up computers and begin firing up lasers. 7 pinholes, 7 lasers, each laser is assigned a respective pinhole which is obvious in the software, first comes the 488nm, then the 640nm, 405nm follows soon after and then comes 355nm. Brent warns us before the UV is lit-up and with good warning as a business card quickly begins to smoke as its dipped into the light path. 532nm and 561nm sit smartly side-by-side and follow each other through the pinholes to the intercept point on the stream, 532nm light is rejected using a 530/40 rejection band with a width of 2nm and we soon learn about alternative options for optimal FITC detection.
We install the 86um tip, running at 37psi, 59kHz, and set up a stream to sort the drop at a delay of roughly 40.1, the camera sits steady at 231, the sorter is ready. 6 way stream separation is made possible with clever modifications and plate zapping is eliminated with the use of conductive tape, the stream is steady as a rock and the droplet formation looks solid, consistent and clean. 6 streams sit in test mode as Brent aligns the collection tubes using the motor driver (unlike old Vantage manual stream movements). 6 peak beads are sorted into each collection tube and each pass above 99% purity with ease, installation is complete.
So many similarities to our old Vantage sorter, so many modifications and so many improvements makes this transition from Vantage sorting to Influx even sweeter, the day ends with a quick lesson in wet and dry shutdowns and we look forward to running samples tomorrow, keen to see 6 way sorting of cells and work out sort modes - excited and ready for day 3.
Chris
