Laboratory of cell and tissue cultures 

BD FACSAria characteristics

Cell phenotyping

• The instrument can be installed and operated in virtually any room without special facility requirements
• The cuvette flow cell achieves high sensitivity in signal detection with low-powered lasers, allowing air-cooled and solid state lasers to be utilized in this system. This, in turn, eliminates the need for special power and cooling requirements associated with high-powered lasers used in conventional stream-in-air sorters.
• The fixed optical alignment frees the operator from tedious instrument optimization, allowing the focus to center on the science rather than the instrument
  
• Because the placement of the sample core stream within the cuvette flow cell and the laser alignment are fixed, daily instrument optimization of the cytometer is no longer necessary.
• A gel-coupling cuvette flow cell collects at least four times the amount of light over stream-in-air sorters: sensitivity in signal detection <125 Molecules of Equivalent Soluble Fluorescein [MESF]
• The collection optics are engineered to achieve the greatest amount of signal detection from each laser. Since reflection is more efficient than transmittance, this design greatly increases the multicolor detection capabilities of the instrument
  
• A fluidics cart allows the instrument to operate in any lab or room without the special and costly facility requirements of a traditional high-speed flow cytometer
• The instrument has a specialized sample injection chamber that accommodates different tube sizes to provide flexibility in experimental design
• After each tube is removed, sheath fluid flushes the sample tubing inside and out to eliminate potential sample carryover. Using a special cleaning mode, the entire sample injection chamber can be filled with ethanol
• The electronics simplify compensation of all fluorescent signals with no limitation of inter- and intra-beam compensation
• Faster signal processing enables acquisition rates of up to 70,000 events per second for 15 parameters
• BD FACSAria™ software provides unparalleled instrument control for flexible sorting and data analysis
• The clog detection feature signals the instrument to stop the sort and protect the collection tubes if the instrument cannot maintain the stream breakoff
• Coefficient of variation PI-Area of <3.0%, full G0/G1 peak for propidium iodide (PI)-stained chicken erythrocyte nuclei
• Sensitivity enables separation of fixed platelets from noise, identification of bacteria, and 0.5-micron beads
• Forward and side scatter performance is optimized for resolving lymphocytes, monocytes, and granulocytes
• Maximum acquisition rate: 70,000 events per second with 8 parameters, 12 compensation pairs. Maximum rate < 70,000 events per second with more parameters, > 100,000 events per second with fewer parameters
  
• Height, Area, and Width measurements available for any parameter. Ratio measurements are also available
• BD FACSDivaTM software with Data File Structure Flow Cytometry Standard (FCS) 3.0 or 2.0, export data to MS Word, MS Ехсеl та Adobe PDF

Cell sorting

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• Due to the unique design of the cuvette flow cell, drop drive disturbances that create noise on stream-in-air systems are not seen on the BD FACSAria cell sorter
• Unlike stream-in-air systems, there is no perceptible performance degradation of any parameter while using the drop drive. Thus, data quality is not compromised during sorting
• Fixed nozzle alignment ensures the stream will return to the same spot after a nozzle tip is removed and reattached
• The 70- and 100-micron nozzle tip sizes are designed to accommodate most cell types. The nozzle sizes enable sorting at a variety of pressures and speeds
• Two- and four-way sorting are standard features available for a variety of bulk collection devices to accommodate different tube sizes (for example, microtubes, 12 X 75 mm, and 15 mL)
• Automatic Cell Deposition Unit (ACDU) option allows for slide and plate sorting into 6, 24, 48, 96, and 384 plates
• The tube holder agitates the sample, keeping cells constantly suspended throughout a long sort
• Advanced digital electronics improve sort performance and provide significant advantages in instrument ease-of use
• The electronics eliminate dead time. This improves the number of signals that can be processed and sorted, thus increasing sort yield by reducing the number of electronic aborts
• Integrated BD FACS™ AccuDrop technology allows the user to determine an accurate drop delay value
• Water recirculator for refrigeration/heating
• Clean flow cell
• Clean sample injection chamber
• Aseptic Sort Setup mode
  
• Sample input, software-adjustable: 4, 20, 37, and 42 degrees C
• Increased drop resolution provides accurate investigation of each sorted drop and results in better sort purity and yield
• At 70 psi and 90 kHz with an average threshold rate of 25,000 events per second for a four-way sort results in a purity of > 98% and a yield > 80% of Poisson’s expected yield
• Sorts were performed at a variety of sheath pressures using cell lines and human peripheral blood mononuclear cells. All sorts resulted in cells that proved viable and proliferated for several days post-sort