Difference between revisions of "Team:SJTU-BioX-Shanghai/Results"

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             </h2>
 
             </h2>
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             <p>xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxxxxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxxxxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx
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             <p>爬片
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A series of experiments were done to determine whether the cell surface display system(CSDS) correctly transports the antibody Ts to the outer membrane of the bacterium, as well as whether the antibody Ts are effective to build a cell-bacteria junction. We used cell-climbing cover glasses as the solid basement on which the cell-bacteria adhesion experiment was carried out. These cover glasses were carefully observed via the fluorescent microscope after a period of incubation, and in what pattern the bacteria adheres to the cell could be determined. 
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()
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流式T抗原
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Certificate the expression of HT29 surface T antigen
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A specified antibody against T antigen was applied to detect the antigen on HT29’s surface.We used FACS to analysis the antigen-antibody reaction. The antibody PNA
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流式粘附
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Quantitative Analysis of the Validity of Antibody Ts
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Next, we were able to quantitatively determine whether the cell surface display system(CSDS) transports the antibody Ts to the outer membrane of the bacterium correctly, and how the antibody works. the FACS analysis was applied to the HT29 cell suspension incubated with antibody-expressing BL21. In this case, BL21 with antibody displayed on the outer membrane should attach to the HT29 cell which correspondingly have the T antigens on its surface. The reaction system was treated with 7AAD by which the dead cells can be differentiated from living ones in a low dye concentration after the co-incubation1. And the BL21 expressed EGFP in order to be detected.
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To identify the positive signal, that is, the bacteria adhered to the HT29 cell, from the cell-bacteria suspension, we decided to screen out the free BL21 before we exam the fluorescence intensity. The signal that shows a similar scale of cells as well as strong fluorescence intensity is inferred as the positive signal after free bacteria are excluded(Fig.X) . In order to draw a statistically meaningful conclusion, each group had a four-times parallel repeat. As shown, positive rate in INP-EGFP(BL21 with INP and EGFP expression) was substantially higher than that of EGFP(BL21 with only EGFP expression) group(Fig.X+1).
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Revision as of 07:35, 14 October 2018

Results

Section1

Instructions: you have the change not only the main text, but also have to modify the text in the navgation bar on the left side. Now put your content here and do the same for the following sections. xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx Note template is here --- OD OD Optical density .

Section2

爬片 A series of experiments were done to determine whether the cell surface display system(CSDS) correctly transports the antibody Ts to the outer membrane of the bacterium, as well as whether the antibody Ts are effective to build a cell-bacteria junction. We used cell-climbing cover glasses as the solid basement on which the cell-bacteria adhesion experiment was carried out. These cover glasses were carefully observed via the fluorescent microscope after a period of incubation, and in what pattern the bacteria adheres to the cell could be determined. () 流式T抗原 Certificate the expression of HT29 surface T antigen A specified antibody against T antigen was applied to detect the antigen on HT29’s surface.We used FACS to analysis the antigen-antibody reaction. The antibody PNA 流式粘附 Quantitative Analysis of the Validity of Antibody Ts Next, we were able to quantitatively determine whether the cell surface display system(CSDS) transports the antibody Ts to the outer membrane of the bacterium correctly, and how the antibody works. the FACS analysis was applied to the HT29 cell suspension incubated with antibody-expressing BL21. In this case, BL21 with antibody displayed on the outer membrane should attach to the HT29 cell which correspondingly have the T antigens on its surface. The reaction system was treated with 7AAD by which the dead cells can be differentiated from living ones in a low dye concentration after the co-incubation1. And the BL21 expressed EGFP in order to be detected. To identify the positive signal, that is, the bacteria adhered to the HT29 cell, from the cell-bacteria suspension, we decided to screen out the free BL21 before we exam the fluorescence intensity. The signal that shows a similar scale of cells as well as strong fluorescence intensity is inferred as the positive signal after free bacteria are excluded(Fig.X) . In order to draw a statistically meaningful conclusion, each group had a four-times parallel repeat. As shown, positive rate in INP-EGFP(BL21 with INP and EGFP expression) was substantially higher than that of EGFP(BL21 with only EGFP expression) group(Fig.X+1).

Animal experiment

Overall workflow of animal experiment

2018 Interlab Plate Reader Protocol
Protocols/Transformation

section4

xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx The figure template is here.

Fig 1. The particle standard curve obtained form the 2nd calibration experiment.

xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx.

The table template is here.

Table 1. Colony forming units per 0.1 OD600

samples dilution factor CFU/mL
8×104 8×105 8×106
1.1 TNTC 48 11 3.84E+07
1.2 248 41 10 3.28E+07
1.3 172 54 5 4.32E+07
2.1 TNTC 143 20 1.14E+08
2.2 TNTC 153 25 1.22E+08
2.3 TNTC 151 18 1.21E+08
3.1 TNTC 119 16 9.52E+07
3.2 TNTC 125 19 1.00E+08
3.3 TNTC 89 18 7.12E+07
4.1 TNTC 209 16 1.67E+08
4.2 TNTC 130 17 1.04E+08
4.3 TNTC 164 10 1.31E+08

Section5

XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX XXX