BD FACSAria Quick Reference Guide
Use this quick reference guide once you are familiar with the procedures in the BD FACSAria User’s Guide .
Starting Up the Cytometer
Follow these steps to start up your BD FACSAria™ cytometer. For more details, refer to Chapter 4 in the user’s guide.
1Start up the workstation.
2Open the flow cell access door to block the lasers.
3Turn on the cytometer main power and the lasers. Wait 30 minutes for
the lasers to warm up.
;Tip    If you are using temperature
control, start heating or cooling the sample injection chamber or the recirculating water bath while the lasers are warming up.
4Launch BD FACSDiva™ software and log in with your user
name and password.
5Verify fluidics levels in the Cytometer window. Replenish
fluids or empty the waste, if needed.
Do not allow the lasers to contact the cuvette flow cell before you activate the stream. Running the lasers without activating the stream
can degrade the performance of the cuvette flow cell.
main power
sheath waste bleach DI ethanol
6Choose Cytometer > Fluidics Startup and click OK to run startup.7Verify the nozzle size, then use s
oftware controls to start the stream.
As a general guideline, the nozzle opening should be at least three times the diameter of the particle to be sorted. If necessary, change the nozzle before you start the stream.
•Click the Sorting button () on the Workspace toolbar to display the Breakoff and Side Stream windows.
•Click the Stream button () in the Breakoff window to turn on the
stream.8Check the stream position in the waste aspirator and inspect the stream
image in the Breakoff window.
If you see any dripping or spraying or the stream appears abnormal, turn off the stream and refer to Troubleshooting the Stream in Chapter 7 in the user’s guide.
9Choose a new Sort Setup mode, if necessary.
10Adjust the Amplitude to achieve fast-merging satellites within 6 drops.
For troubleshooting assistance, see Troubleshooting the Breakoff on page 5 or refer to Sorting Troubleshooting in Chapter 7 in the user’s guide.
11Enter the generated value as the Drop 1 target.
12Turn on the Sweet Spot when the drop pattern is stable.
Sort Setup Nozzle (μm)
Recommended Applications
High 70High-speed, high-throughput sorting Medium 70Sorting of cell lines, fragile cells
Low 100Single-cell sorting, ACDU sorting, sorting of large or fragile cells
degrade
Custom
70
Non-sorting mode for high sensitivity
Sorting Setup
Do the following to set up for sorting. For more details, refer to Chapter5 in the user’s guide.
1Check the laser delay and area scaling factors for your sheath pressure.
Refer to Checking Performance with Cytometer Setup and Tracking in Chapter 4 in the user’s guide.
2Optimize cytometer settings for the sample to be sorted.
Refer to Data Collection in Chapter 4 in the user’s guide.
3Install the required collection device and set up the side streams.
4Use BD FACS™ Accudrop to calculate the drop delay.
5Run the sample to be sorted. Use gating tools and subsetting methods to define the population(s) of interest.
NOTICE Snap-to gates cannot be used for sort gates.
6Define a Sort Layout for the tube containing the defined sort populations.
Sorting
1Install the collection tubes, plate, or slide.
2Install the sample tube on the loading port and click Load.
3(Optional) Turn on the deflection plates and open the aspirator drawer. 4Verify that the current tube pointer is set to the appropriate tube in the Browser and click Sort.
5Click OK if you are prompted to open the aspirator drawer or turn on the deflection plates.
Sorting continues until the required number of cells has been sorted. If the number of Target Events is set to Continuous, sorting continues until you manually stop sorting by clicking the Stop Acquiring or Sort button.
;Tip    Click Record Data to save data for the tube. Acquisition and sorting continue when the required number of events has been recorded. Shutting Down the Cytometer
1Unload your sample tube, if one is loaded.
2Open the flow cell access door to block the lasers.
3Turn off the stream.
4Run fluidics shutdown.
5Follow the prompts on the screen to shut down the fluidics system.
6Turn off the cytometer main power, quit BD FACSDiva software, and shut down the computer.
7(Optional) Remove the nozzle and allow it to air dry.
Troubleshooting the Breakoff
A
b n o r m a l  S t r e a m  I m a g e
Normal stream image P o s s i b l e C a u s e s
Nozzle i nserted improperly
Nozzle i nserted improperly or orifice off center Partial clog
Wet or dirty strobe lens
Attenuation on at the wrong pressure R e c o m m e n d e d S o l u t i o n s
Remove the nozzle and re-insert it.
Remove the nozzle, adjust the opening, and re-insert it.
Remove the
nozzle, clean it, and re-insert it.Clean the lens as described in the user’s
guide.
Turn off attenuation in the Side Stream window.
bdbiosciences
BD Biosciences 2350 Qume Drive
San Jose, CA 95131-1807USA
Tel (877) 232-8995Fax (800) 325-9637Brazil
Tel (55) 11-5185-9995Fax (55) 11-5185-9895
Europe
Tel (32) 2 400 98 95Fax (32) 2 401 70 94
Japan
Nippon Becton Dickinson Company, Ltd.Toll Free 0120-8555-90Asia Pacific
Fax (65) 6-860-1590
Canada
Toll Free (888) 259-0187Tel (905) 542-8028Tel (65) 6-861-0633*****************
**************************
Tel 81-24-593-5405Fax 81-24-593-5761
Mexico
Fax (888) 229-9918*************Toll Free 01-800-236-2543Tel (52) 55 5999 8296Fax (52) 55 5999 8288
BD
FACSDiva Software Toolbars
Workspace toolbar
Browser toolbar
Save Browser
Cytometer Worksheet Biexp. Editor Inspector
Acq.Sorting
New Folder
New Global
New Expt
New New Tube
New Cytom. New Sort Settings Layout
Worksheets
View Select
Dot
Plot
Contour
Plot Histogram
Worksheet toolbar
Zoom In Zoom Out
Increase Plot Size Decrease Plot Size
Autopolygon Gate Snap-To Gate Polygon Gate Rectangle Gate Quadrant Gate
Interval Gate Autointerval
Gate Snap-To
Interval Gate
Arrow Line
Text
Align Left Align
Right
Align Top
Align Bottom
Distribute Horizontally Distribute Vertically
Make Same
Size
Dashboard
Worksheet Save as
PDF
Print Show Grid
Snap-To Grid
Specimen

版权声明：本站内容均来自互联网，仅供演示用，请勿用于商业和其他非法用途。如果侵犯了您的权益请与我们联系QQ:729038198，我们将在24小时内删除。