Luminex Vendors Wooing New Customers

Is it just me, or have you been inundated with vendors trying to get you to use their bead-based assay kits. The two big wigs in this field are BioRad and Millipore (listed alphabetically as to not show any preferential treatment on my part) and they really want your business. As you may know, the Flow Cytometry Facility has a BioPlex machine in the Kovler Facility which gets moderate use on a relatively regular basis. I've met with both companies' sales force and they are very interested in hosting some seminars to show their products' worth. Being an equal opportunity obliger, I agreed and so we will be hosting 2 seminars regarding multiplex analyte kits. The first one will be from BioRad on February 26th at noon from 10:30 to 11:30 in Cummings Room 119. It will focus mostly on signaling protein detection from cell lysates, but I'm sure the technology in general will be discussed. This one will have food too! The second one from Millipore will be held on March 12th at 9AM also in Cummings Room 119 (go figure). The Millipore people are also bringing in some kits for you to test out some samples on the 12th and 13th, so this is a great opportunity to try something for free. If you need any information on this, send an email to ucflow'at'gmail'dot'com

Aria to Aria II upgrade...Painful, but worth it

Well that took way longer than expected. A two and a half day estimate went into its 5th day last friday. If anything could have gone wrong, it did. The procedure was going really well for the first two days, and then came the software. Why is it that whenever a problem occurs it always involves software glitches? Needless to say, it was a very frustrating afternoon on Thursday when we were completely stuck until the software started working properly. But, after some hard work and perseverance we finally got things up and running. We were able to start sorting on Friday, and to tell the truth, once we started, it became clear why we went through the trouble in the first place. The Aria started up so smoothly, and ran without a hiccup. You really can just pop in the nozzle and bam, the droplets just fall right into position. If you've ever tried to put the old-style nozzles in an Aria I (or had to use the wretched gold plated nozzles), then you'll really appreciate how easy it is on the Aria II. We've been running mostly with the 85um tip and have come to really appreciate that size. The 70um tip is good for going fast, but if the sample prep is not perfect, and the sample size homogenous and small, you get small jumps in the side streams which can really affect purity and yield. The 85um tip provides just enough room to still go pretty fast, and it's able to handle some junk too. The other thing that is instantly evident once you walk into the room is just how quiet the instrument is. The Aria I sounded like a rocket was taking off constantly, but now it's so quiet it's almost a little erie. Lastly, the charge plates are really nice. 4-way sorting is not even an issue any longer. In fact, the voltage sometimes is a bit too high so we've had to lower it to avoid hitting the plates with the side streams. So far, we've been really impressed with the performance of the instrument. One thing we're eager to get a feel for is how easy it is to keep clean. The jury is still out on that one, but we'll keep a close eye on it for sure.

Aria to Aria II upgrade...Day 2

Getting closer! Much of the Fluidics Board is hooked up and ready to go, the internal fluidic lines have been swapped. Today we'll tackle the flow cell, sample chamber, and sort block, and hopefully later, software and alignment. It probably seems like nothing has been done yet, but let me tell you, lots has been done. Bill and Scott (BD Engineers) have filled a 3' x 3' x 1.5' box with parts from the Aria I. Which reminds me, we'll have to find a new home for some of these parts, and the rest of them, well looks like I'll have to dig through some old Make magazines and see what we can put together.

Aria to Aria II upgrade...Day 1

The Flow Lab's legacy FACSAria is finally getting its fluidics upgrade today. The process takes about 3 days, so I'll give you some updates via photos. Day 1 was basically demo day, and the pic displayed here is the Fluidics area inside which has been totally ripped out. It looks a bit messy now, but the new system is much more stream-lined than the old. For those of you not familiar with the benefits of the fluidics upgrade, I'll give you a quick run-down and a bit of history. 1st the history. Sorters, for years have used a very simple process for driving fluidics; fill a tank with PBS, pressurize the tank at constant pressure, which causes the fluid to move through the system at a constant velocity, thereby giving a stable droplet break-off pattern. Enter BD and their quest to put a sorter on every researcher's desk. BD developed a fluidics system which was marketed as a "bench-top" sorter which could be run by anyone. Everything is automated, cleaning, sterilizing, running, monitoring, etc... In attempting to make this "easy-to-use" sorter, they created a service nightmare. There were so many valves and switches and pumps put in to do the simple job of moving PBS from a tank to a flow cell that at any given point, one of them was not working properly. Additionally, with all the valves and switches and automation came many small pieces of tubing and hangouts for bugs and bacteria. This created a situation where we had to jump through hoops trying to keep the sorter clean. Other sorters, like the MoFlo still had the classic fluidics system and, at least in our hands, had almost zero fluidics problems over about a 10 year period, and very few contamination issues with little effort. After a few years of complaints and retooling, BD releases the FACSAria II, and guess what? They got rid of a ton of valves, switches, hideouts, and once again went back to a simplified, robust fluidics system. It still has a few bells and whistles, but certainly not as grotesquely over-engineered as the Aria I. Will it withstand the onslaught of abuse only a core facility like ours can dole out? Only time will tell. Regardless, you'll hear about it here, so stay tuned.

How to choose - Part I

I am constantly reviewing new flow cytometry related technology, and I must admit, it's a pretty fun part of the job. So, over the years I think I've developed some strategies for choosing the proper piece of equipment which I will attempt to share in this "How to Choose" series. The Flow Cytometry Facility was recently awarded a Shared Instrumentation Grant from the NIH to purchase the LSRII it has been leasing for the past 8 months (see previous post for details). In approaching the original plan to acquire a new high-end analyzer, I went through this time-consuming process of evaluating multiple pieces of equipment in order to decide which one to get. Therefore, I will attempt to outline some of the key aspects which can hopefully translate to technology and equipment outside of flow cytometry. In general, the steps for choosing the proper instrument include, 1. Query your user base, 2. Establish selection criteria, 3. Research available technologies, 4. Match available technology with criteria, 5. Weigh non-technical aspects of ownership (e.g. Service, Infrastructure Necessities, Space, etc.), and 6. Test the instrument with real samples. I've gone through this process for equipment as complex as cell sorters, and as simple as incubators, and every time I feel the proper decision was made. The next few postings will detail the specifics of each of these 6 steps, so be sure to check back soon for an update.

Flow Cytometry Facility awarded NIH SIG

The Flow Cytometry Facility is pleased to announce that we have been awarded a Shared Instrumentation Grant from the NIH, which will be used to pay off the LSRII-Blue we have been leasing since April 2008. The facility recognized the necessity for such a high-end benchtop analyzer that we proceeded to lease the LSRII while resubmitting the grant proposal for the instrument. Needless to say we are very happy with the outcome of this proposal, and with the performance of the LSRII thus far. When publishing data using the facility's LSRII, you may acknowledge the facility as well as the SIG grant number for this instrument, 1S10RR24570-01A1.

"Lasers"

Do you like Star Wars? I do. I'm not one of those freaky, dress-up like Chewbacca fans, and talk like yoda, I do not, but what I really like about Star Wars are the lasers. It just seems like such a practical way to shoot things. No need to carry along a bunch of ammunition, all you need is your trusty multi-KiloWatt laser. Another benefit is that light travels, well, at the speed of light, much faster than a speeding bullet or a falling bomb. Lasers are used all over the place, most-notably for me, Flow Cytometry and Microscopy, but now it seems like the vision of Star Wars may be closer than ever. Darpa (The advanced research division of the Department of Defense) has been investigating weapons-grade lasers for some time, but with the use of some newer technologies, there may actually be a viable option. HELLADS, or High Energy Liquid Laser Area Defense System, a laser system developed by Textron, is getting some big government bucks to design and test a 150kW Laser Weapon System that could be carried on a Fighter Jet and be used to shoot down missiles. So, what's so special about the laser? It seems this laser system is using a series of thin ceramic slabs, bathed in rapidly circulating coolant to achieve a high power output while maintaining a non-destructive temperature load. These so-called slab lasers are attempting to tackle a long-standing problem with lasers, what to do with waste heat. If successfully employed, these types of technologies may trickle their way down to us lowly folks in the flow cytometry world, and I may actually be able to realize my goal of a centralized laser depot with a bunch of fiber-optic cables extending out of it. I've already got a name for it...Medussa! If you have a high enough laser source, you could split that beam a bunch of times, and using fiber optic cables, bring the light to any number of instruments using a "plug-n-play" connection scheme. Some day...