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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.
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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
II FlowJo [email protected] 400-680-5527
FlowJo ....................................................................................................47 .........................................................................................................................47 FlowJo .................................................................................................48 .................................................................................................51 .........................................................................................................................53
PatrickUnderline
III FlowJo [email protected] 400-680-5527
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
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
3 FlowJo [email protected] 400-680-5527
NEXT
NEXT
Install
Done
4 FlowJo [email protected] 400-680-5527
a. DongleFlowJo
b. Serial NumberUse
Serial Number
c. Get Trial LicenseFlowJo
Use Serial Number
5 FlowJo [email protected] 400-680-5527
d. DemoFlowJo
Run in Demo ModeDemoFlowJoDemo
FlowJo
FlowJo
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
7 FlowJo [email protected] 400-680-5527
3 color comp3 color
comp
Cy5PE comp.fcs
8 FlowJo [email protected] 400-680-5527
1) XSSC
SSC
2) YFSC
FSC
3)
:
9 FlowJo [email protected] 400-680-5527
1) XCy5PE:CD4YHistogram
2)
a.
CD4CD4
10 FlowJo [email protected] 400-680-5527
b.
CD4CD
a.
WORDPPT
b.
SVG
PNGEMFJPG
11 FlowJo [email protected] 400-680-5527
c. FlowJo
d.
12 FlowJo [email protected] 400-680-5527
e.
data001.JPGJPG, PNG, GIF, EMF,
SVG, PDF
FlowJoPNG
f.
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
14 FlowJo [email protected] 400-680-5527
3. XFluor::CD3YFSC
CD3T
15 FlowJo [email protected] 400-680-5527
4. TXCy5PE::CD4Y
PhyEry::CD8
Q1CD8CD8+ T
Q2
Q3CD4CD4+ T
Q4
16 FlowJo [email protected] 400-680-5527
17 FlowJo [email protected] 400-680-5527
TREGCD4/CD25/FOXP3SSC vs. CD4
CD4+CD25 vs. FOXP3
CD25+/FOXP3+CD4+/CD25+/FOXP3+
TREG
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
19 FlowJo [email protected] 400-680-5527
2.
Sample
FlourFITC comp.fcs
20 FlowJo [email protected] 400-680-5527
3. 23FlowJo
4.
>
21 FlowJo [email protected] 400-680-5527
(poor control )FlowJo
1. 1,2,3
FlowJoPE
22 FlowJo [email protected] 400-680-5527
1) PEcw_Size
2) Clear
2.
1) PEFSC/SSC
23 FlowJo [email protected] 400-680-5527
2) XPhyEry::aCD8Y
Histogram PEPE
a.
b.
3) PE-PE
PE+
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%
25 FlowJo [email protected] 400-680-5527
2. XYT
3. T
Change transform values
26 FlowJo [email protected] 400-680-5527
X-10-30
a.
b. FCS3.0FlowJoFCS2.0
FlowJo
27 FlowJo [email protected] 400-680-5527
FlowJo
Titration DataCD8Sample 1
Sample 2 ~ Sample 82
1.
Sample 1
28 FlowJo [email protected] 400-680-5527
XFL1-HYHistogram Sample 1
FL1-H+
RenameCD8+FL1-H-
CD8-
2.
29 FlowJo [email protected] 400-680-5527
CD8+
MedianParameter
FL1-HCD8FL1-H
30 FlowJo [email protected] 400-680-5527
(FL1-H):MedianCD8-CD8
FL1-H
3.
Sample 1Sample 2
Sample 2Sample
1
31 FlowJo [email protected] 400-680-5527
Sample 1
Titration Data
Sample 1
ALTSample 1
Titration Data
Sample 1
32 FlowJo [email protected] 400-680-5527
1.
Sample 1CD8+CD8-
(FL1-H):Median
33 FlowJo [email protected] 400-680-5527
2.
Keyword
Antibody Concentration
OK
3.
CD8+ (FL1-H) Median CD8- (FL1-H) MedianCD8
CD8MFI
OK
34 FlowJo [email protected] 400-680-5527
35 FlowJo [email protected] 400-680-5527
4.
MFI
1MFI1.
1.
Sample 1
36 FlowJo [email protected] 400-680-5527
FlowJoTitration Data
FlowJo
37 FlowJo [email protected] 400-680-5527
2. (Overlay)
1
Titration Data
(1) Sample 1Sample 2
Sample 1
(2) SpecifyControl
Sample 1Sample 1OK
38 FlowJo [email protected] 400-680-5527
(3)
39 FlowJo [email protected] 400-680-5527
2
Titration Data
(1) Sample 1CTRL
Sample 1
Sample 1
40 FlowJo [email protected] 400-680-5527
(2)
41 FlowJo [email protected] 400-680-5527
(3)
(4) coloringfilled
(5)
42 FlowJo [email protected] 400-680-5527
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
44 FlowJo [email protected] 400-680-5527
FlowJo
apoptosisZL-2011-12-20_400.fcs
1. FlowJo
XFSC-A,YSSC-A
2. SSC/FSC
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-
46 FlowJo [email protected] 400-680-5527
Q2+Q3
AnnexinV
47 FlowJo [email protected] 400-680-5527
FlowJo
G1
SG2MG1S
DNAG2DNA
M
DNAPIPI
DNARNA
DNA
DNA
48 FlowJo [email protected] 400-680-5527
FlowJo
cell cycle 5206.001PI
1.
5206.001XFSC-HY
SSC-H
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
50 FlowJo [email protected] 400-680-5527
3. FlowJo
G1SG2G1CV
G2CV
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
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
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