Below are several tools, procedures, tips-and-tricks that may be handy for flow operators.
Rmax is a fast and simple method to assess instrument or sorter performance (Riddell et al., 2015). This method was developed by Andy Riddell and Alexis Gonzalez at EMBL in collaboration with our lab and Lola Martinez at CNIO. Rmax is defined as the maximum sort Recovery possible for the current conditions in which the sort is taking place, and it is determined exclusively by measuring the frequency of the target population in the original sample and the frequency of unsorted target population found in the centre stream catch.
This is an ideal method to measure how good the sorter is at charging the right drop at the right time. This can be performed with an ideal sample based only on beads avoiding wasting precious sample (see video on the right). In addition, sort performance can be systematically compared for different sorting conditions (sort decision, drop drive frequency, nozzle size, etc), to control daily drop delay determination, and to troubleshoot the instrument. More information can be found below:
Article - Andrew Riddell, Rui Gardner, Alexis Perez-Gonzalez, Telma Lopes, Lola Martinez (2015) Rmax: A systematic approach to evaluate instrument sort performance using center stream catch, Methods,doi:10.1016/j.ymeth.2015.02.017
Remote access technologies not only allows sharing computer desktops between users, but also to control the connected instrumentation remotely. There are currently many different software packages available. Choice is according to technical requirements, IT permissions, and price.
Our optimized setup uses the equipment and configuration as follows:
Brief tutorial on how to change the sample agitation motor o-ring in the bulk injection chamber of the FACSAria.
Learning to operate a MoFlo High-Speed Cell Sorter follows a steep curve, requiring a few weeks of dedicated training. Some users may, however, need to shutdown the MoFlo on their own. This can be achieved as long as the shutdown procedures are carefully followed, and after going through these procedures with the facility's staff. The complete operating procedures for the MoFlo can be downloaded using the link below, and are intended for cell sorter operators.
Given the large diversity of research areas within the IGC, the need to sort cells ranging from bacteria to mammalian cells can be a challenge.
When sorting bacteria or small microorganisms, the sample's fluidic path must be clear of debris of larger cells. When sorting eukaryotic cells, the sample fluidic path must not be contaminated by bacterial colonies. To avoid buying two cell sorters that would be dedicated to both types of cells, we put together two different fluidic assemblies, that can be setup in the machine in less than 15 min.
Material and assembly instructions:MoFlo Nozzle and Fluidic Assembly
Download a simple procedure to restore FACSDiva Database after Communication with database error.