By nsiddiqui|2017-11-09T20:55:31+00:00November 9th, 2017|Troubleshooting|Comments Off on Droplet containment vacuum not functioning
How many cells do I need to bring to the sorter?
This is the most commonly asked question. To answer it we need to know the following information: What is the approximate percentage of the population(s) you wish to sort? How many total cells do you want back? Do you want stringent purification or enrichment? How fragile or large are your cells? All of these [...]
How long will it take to sort?
The answer to this question depends primarily on your goal (enrichment vs stringent purification), the nature of your cells (fragile cells or large cells need to be sorted at lower pressures and speeds with a larger nozzle), and the concentration of your sample (more dilute samples will take longer to sort). For example, on [...]
Do I need to filter my cells?
It is REQUIRED that your cells be filtered through at 70um nylon mesh, preferably right before running on the sorter. We can provide the mesh if necessary.
What do I re-suspend my cells in for sorting?
Proper sample preparation is absolutely essential for obtaining good purity and yields. After the final wash, cells should be re-suspended at concentrations of 20-30 million (for primary cells) or 5-10 million (for cell lines) per mL in FACS Tubes (see below). The media can be any buffered isotonic salt solution (such as PBS) but [...]
What controls do I need to bring?
You will need to bring an unstained control, single colour compensation controls and you MAY need to bring FMO (Fluorescence Minus One) gating controls. You may want to consider an FMO control if you are using more than 5 colours simultaneously and if your populations of interest cannot be discretely separated from background/negatives. Please [...]
What are FACS Tubes and where do I get them?
BD Falcon makes FACS Tubes of various kinds. For all cell sorters, order 5 ml sterile polypropylene tubes (catalogue number BD2063). For non-sterile and analytical samples order catalogue number BD2002 (polypropylene) or BD2052 (polystyrene).
What kind of tubes or plates should the cells be sorted into?
For abundant populations we recommend 50 mL (MoFlo) or 15 mL (FACS Aria) Falcon Tubes, or 5 mL Falcon tubes. We can also sort into a range of microtiter plates, including from 96-well to 8-well, microscope slides or Petri dishes. Please talk to us about your requirements and we will advise you on the [...]
