flowjo 中文实用手册
DESCRIPTION
the chinese version of Flowjo guide, simple but straight forward, explaning the basic of how to use Flowjo, like gating and backgating, worksheet, table and compensation etc.TRANSCRIPT
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I FlowJo [email protected] 400-680-5527
FlowJo FlowJo7.6.5 2012/06/20
FlowJo ......................................................................................................................................1 FlowJo ...............................................................................................................1 FlowJo(Windows) .........................................................................................................1 FlowJo ...............................................................................................................................5
FlowJo ......................................................................................................................6 ...........................................................................................6 .........................................................................................................................10
FlowJo ............................................................................................................13 .................................................................................................13 .....................................................................................................................16 .........................................................................................................................16
FlowJo ....................................................................................................................18 FlowJo..............................................................................18 FlowJo .........................................................................................................18 .....................................................................................................................................24
FlowJo ....................................................................................................................27 .........................................................................................................................27 .........................................................................................................................32 .........................................................................................................................35
FlowJo ............................................................................................................43 .................................................................................................................................43 FlowJo .........................................................................................................44 .........................................................................................................................45
PatrickUnderline
PatrickUnderline
PatrickUnderline
PatrickUnderline
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II FlowJo [email protected] 400-680-5527
FlowJo ....................................................................................................47 .........................................................................................................................47 FlowJo .................................................................................................48 .................................................................................................51 .........................................................................................................................53
PatrickUnderline
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III FlowJo [email protected] 400-680-5527
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1 FlowJo [email protected] 400-680-5527
FlowJo
FlowJoLeonard HerzenbergFACS
90FlowJo
FlowJo
FlowJo
1. PC
Windows XPVistaWindows7
512MB
2.
OSX10.3
512MB
FlowJo(Windows)
http://www.flowjochina.com/content/download
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2 FlowJo [email protected] 400-680-5527
V7.6.5
http://www.flowjochina.com/sites/default/files/7.6.5_X32.exe
FlowJo
FlowJo
NEXT
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3 FlowJo [email protected] 400-680-5527
NEXT
NEXT
Install
Done
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4 FlowJo [email protected] 400-680-5527
a. DongleFlowJo
b. Serial NumberUse
Serial Number
c. Get Trial LicenseFlowJo
Use Serial Number
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5 FlowJo [email protected] 400-680-5527
d. DemoFlowJo
Run in Demo ModeDemoFlowJoDemo
FlowJo
FlowJo
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6 FlowJo [email protected] 400-680-5527
FlowJo
,flowjochina
www.flowjochina.com/sites/default/files/demodata.rar
3 color compCy5PE comp.fcs
1. FlowJo
2. 3 color compFlowJo
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7 FlowJo [email protected] 400-680-5527
3 color comp3 color
comp
Cy5PE comp.fcs
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8 FlowJo [email protected] 400-680-5527
1) XSSC
SSC
2) YFSC
FSC
3)
:
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9 FlowJo [email protected] 400-680-5527
1) XCy5PE:CD4YHistogram
2)
a.
CD4CD4
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10 FlowJo [email protected] 400-680-5527
b.
CD4CD
a.
WORDPPT
b.
SVG
PNGEMFJPG
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11 FlowJo [email protected] 400-680-5527
c. FlowJo
d.
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e.
data001.JPGJPG, PNG, GIF, EMF,
SVG, PDF
FlowJoPNG
f.
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13 FlowJo [email protected] 400-680-5527
FlowJo
FlowJo
FlowJo3-color-experiment931115-
B02-Sample01.FCS
1. 3-color-experimentFlowJo
2. 931115-B02-Sample01.FCS
XFSCYSSC
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14 FlowJo [email protected] 400-680-5527
3. XFluor::CD3YFSC
CD3T
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15 FlowJo [email protected] 400-680-5527
4. TXCy5PE::CD4Y
PhyEry::CD8
Q1CD8CD8+ T
Q2
Q3CD4CD4+ T
Q4
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16 FlowJo [email protected] 400-680-5527
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17 FlowJo [email protected] 400-680-5527
TREGCD4/CD25/FOXP3SSC vs. CD4
CD4+CD25 vs. FOXP3
CD25+/FOXP3+CD4+/CD25+/FOXP3+
TREG
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18 FlowJo [email protected] 400-680-5527
FlowJo
FlowJo
FlowJo
1.
2.
3.
FlowJo
FlowJoFlowJo
4
http://www.video.flowjochina.com/v75%20Compensation_conv.html
1. FlowJo
3 color compCy5PE comp.fcs, FITC
comp.fcs, PE comp.fcs3-COLOR.FCS
Cy5PE, FITC, PE
PatrickHighlight
PatrickHighlight
PatrickHighlight
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19 FlowJo [email protected] 400-680-5527
2.
Sample
FlourFITC comp.fcs
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20 FlowJo [email protected] 400-680-5527
3. 23FlowJo
4.
>
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21 FlowJo [email protected] 400-680-5527
(poor control )FlowJo
1. 1,2,3
FlowJoPE
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22 FlowJo [email protected] 400-680-5527
1) PEcw_Size
2) Clear
2.
1) PEFSC/SSC
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23 FlowJo [email protected] 400-680-5527
2) XPhyEry::aCD8Y
Histogram PEPE
a.
b.
3) PE-PE
PE+
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24 FlowJo [email protected] 400-680-5527
3. FlowJo
3-color comp
>
0Log
0
LogicalFlowJo
3-color comp3-
COLOR.FCS
1. 3-COLOR
XCD3YCD8
gate135.1%
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25 FlowJo [email protected] 400-680-5527
2. XYT
3. T
Change transform values
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26 FlowJo [email protected] 400-680-5527
X-10-30
a.
b. FCS3.0FlowJoFCS2.0
FlowJo
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27 FlowJo [email protected] 400-680-5527
FlowJo
Titration DataCD8Sample 1
Sample 2 ~ Sample 82
1.
Sample 1
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28 FlowJo [email protected] 400-680-5527
XFL1-HYHistogram Sample 1
FL1-H+
RenameCD8+FL1-H-
CD8-
2.
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CD8+
MedianParameter
FL1-HCD8FL1-H
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30 FlowJo [email protected] 400-680-5527
(FL1-H):MedianCD8-CD8
FL1-H
3.
Sample 1Sample 2
Sample 2Sample
1
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Sample 1
Titration Data
Sample 1
ALTSample 1
Titration Data
Sample 1
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32 FlowJo [email protected] 400-680-5527
1.
Sample 1CD8+CD8-
(FL1-H):Median
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33 FlowJo [email protected] 400-680-5527
2.
Keyword
Antibody Concentration
OK
3.
CD8+ (FL1-H) Median CD8- (FL1-H) MedianCD8
CD8MFI
OK
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34 FlowJo [email protected] 400-680-5527
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35 FlowJo [email protected] 400-680-5527
4.
MFI
1MFI1.
1.
Sample 1
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36 FlowJo [email protected] 400-680-5527
FlowJoTitration Data
FlowJo
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37 FlowJo [email protected] 400-680-5527
2. (Overlay)
1
Titration Data
(1) Sample 1Sample 2
Sample 1
(2) SpecifyControl
Sample 1Sample 1OK
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(3)
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39 FlowJo [email protected] 400-680-5527
2
Titration Data
(1) Sample 1CTRL
Sample 1
Sample 1
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40 FlowJo [email protected] 400-680-5527
(2)
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41 FlowJo [email protected] 400-680-5527
(3)
(4) coloringfilled
(5)
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42 FlowJo [email protected] 400-680-5527
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43 FlowJo [email protected] 400-680-5527
FlowJo
Annexin V-FITC/PI
Annexin V-FITC/PI
PSVAnnexin V-FITC
PS
PSPI
PS
AnnexinPI
Annexin V-FITC/PI
AnnexinAnnexin, PI
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44 FlowJo [email protected] 400-680-5527
FlowJo
apoptosisZL-2011-12-20_400.fcs
1. FlowJo
XFSC-A,YSSC-A
2. SSC/FSC
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45 FlowJo [email protected] 400-680-5527
XFITC,AnnexinV-FITCYPE-Texas,PI
1.
2.
Q1(AnnexinV-FITC)-/PI+
Q2: (AnnexinV+FITC)+/PI+
Q3(AnnexinV-FITC)+/PI-
Q4(AnnexinV-FITC)-/PI-
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46 FlowJo [email protected] 400-680-5527
Q2+Q3
AnnexinV
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47 FlowJo [email protected] 400-680-5527
FlowJo
G1
SG2MG1S
DNAG2DNA
M
DNAPIPI
DNARNA
DNA
DNA
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48 FlowJo [email protected] 400-680-5527
FlowJo
cell cycle 5206.001PI
1.
5206.001XFSC-HY
SSC-H
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49 FlowJo [email protected] 400-680-5527
2.
XFL2-W(width)YFL2-A(area)
BDFL2-A
FL2-WFL2-AFL2H
BeckmanXFL3-LIN,PI
YFL3-PEAK,PI
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50 FlowJo [email protected] 400-680-5527
3. FlowJo
G1SG2G1CV
G2CV
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51 FlowJo [email protected] 400-680-5527
http://www.flowjochina.com/content/tutorial-video#cellcycle
1.
RMS
RMS(root mean square error)
RMS3.85
2.
WatsonDean-Jett-Fox
RMSRMS
Could not fit.
Watson
Dean-Jett-Fox
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52 FlowJo [email protected] 400-680-5527
3.
G1G2meanCV
1 G2G12
G1G2G1
2(2G1)G2G1G20.5
2 G1G2
G1mean
G1mean
3 CV
DNAG1G2CV
G1 CV
=G2 CVG2 CV G1 CV
CVCV
CV
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53 FlowJo [email protected] 400-680-5527
1. RMSroot mean square errorRMS
2. G1G1
SS
G2G2
3. G1G1
G2G2
4. CVcoefficient of variationG0/G1
CVCV
G1 cvG1G1
G2 cvG2
Flowjo
FlowJo FlJ FlowJo