BD LSR II 사용자 설명서 - 페이지 4

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BD LSR II에 대해서도 마찬가지입니다: 사용자 설명서 (20 페이지)

c. Repeat for each parameter you're not using

d. Do not delete Parameters that may be of interest.

e. If you add back a deleted parameter to the experiment it will come

in with different voltages, not specified PMT settings by facility.

This may cause problems for resolution. Check cytometer

settings for the 197 bead tube to see correct PMT voltage to

assign when adding back a parameter.

4. Keep forward and side scatter height and width selected

5. The data may be viewed compensated or not, but the uncompensated

data is written to the database along with the matrix so that other

compensations may be used later in FlowJo.

Note: For each tube collected in Diva software, uncompensated data is

recorded and the compensation matrix is applied as a transform, it does not

affect your data.

All samples must be filtered for sorters. The analysis instruments have nylon

mesh squares on the desktop if you need them

Compensation

• Corrects for the emission of one fluor into the detector used to measure another.

• Flow Cytometer use filter sets that are optimized for very specific wavelengths of light.

• Dye/Fluorochrome emissions often span from one filter into another.

• COMPENSATION IS VERY IMPORTANT - without it, our data cannot be interpreted properly.

• To calculate compensation, every experiment needs an unstained control as well as each

fluorochrome singularly.

• Antibody capture beads can be substituted for cell controls.

1. Create Compensation Controls. Select Experiment à

Compensation Setup à Create Compensation Controls.

2. In the Create Compensation Controls

window, ensure that all of your colors are

labeled "Generic". Then select "OK". Do

not compare negative cells to positive

beads or vice versa.