Revision as of 09:11, 25 July 2018

Interlab Study

It is a good opportunity for our team to participate in the project which aims to improve the measurement tools available to both the iGEM community and the synthetic biology community as a whole. And we are glad to cooperate with many other teams from around the world to solve the same problem in synthetic biology. This year's iGEM InterLab study is about could we use CFU/ml to replace the absorbance in fluorescence/od measurement.

To accomplish the goals, we use measure the fluorescence and absorbance with SpectraMax i3x plate reader. In additionally we need to get the competent DH5α and transforme the plasmids into cells. And this wiki will present the workflow and the results we get. We separate the wiki into 3 segments , and the first part is transformation of the plasmids into DH5α.

Transformation

According to the requirements of iGEM, we transformed all the parts (BBa_R0040, BBa_I20270, BBa_J364000, BBa_J364001, BBa_J364002, BBa_J364007, BBa_J364008, BBa_J364009) into competent DH5α. And we successfully get sufficient colonies in all the plates. Therefore we select 2 colonies in each plate to cultivate them In LB medium containing chloramphenicol for overnight with 37℃, 220rpm.

Calibration 1: OD600 Reference point - LUDOX

Through this calibration, we could transform the absorbance (Abs600 ) data from plate reader into a comparable OD 600 measurement in the spectrophotometer, which will help a lot during the following experiments. By doing so, we get the data following.

Table 1: We get the ratiometric conversion factor of 3.549 in our plate reader after measuring the wells adding 100ul Ludox and water.

	LUDOX CL-X	H2O
Replicate 1	0.056	0.039
Replicate 2	0.059	0.039
Replicate 3	0.058	0.040
Replicate 4	0.056	0.040
Arith. Mean	0.057	0.040
Corrected Abs600	0.018
Reference OD600	0.063
OD600/Abs600	3.549

Calibration 2: Particle Standard Curve - Microsphere

Because the monodisperse silica microspheres are similar to the cells in size and optical characteristics during measuring the absorbance in 600nm. It is necessary to construct the particle standard curve and in this way we could change the absorbance into estimated cells.

Table 2 :the raw data we get after diluting a series of monodisperse silica microspheres in the 96 cell well plates.

Number of Particles	2.35E+08	1.18E+08	5.88E+07	2.94E+07	1.47E+07	7.35E+06	3.68E+06	1.84E+06	9.19E+05	4.60E+05	2.30E+05	0
Replicate 1	0.542	0.347	0.202	0.124	0.080	0.059	0.050	0.047	0.043	0.041	0.040	0.042
Replicate 2	0.531	0.344	0.187	0.116	0.077	0.057	0.048	0.042	0.041	0.041	0.040	0.038
Replicate 3	0.794	0.319	0.177	0.105	0.076	0.062	0.049	0.046	0.043	0.042	0.040	0.039
Replicate 4	0.609	0.314	0.193	0.106	0.076	0.057	0.048	0.046	0.043	0.042	0.040	0.042
Arith. Mean	0.619	0.331	0.190	0.113	0.077	0.059	0.049	0.045	0.043	0.042	0.040	0.040
Arith. Std.Dev.	0.122	0.017	0.011	0.009	0.002	0.002	0.001	0.002	0.001	0.001	0.000	0.002
Arith. Net Mean	0.579	0.291	0.150	0.073	0.037	0.019	0.009	0.005	0.002	0.001	0.000

Figure 1：The Particle Standard Curve with monodisperse silica microspheres (log scale)

Figure 2 : The Particle Standard Curve with monodisperse silica microspheres (line scale)

Table 3: The raw data after measuring the fluorescence (excitation light: 485 nm, emission light 530nm) in plate reader.

Fluorescein uM

10.00

5

2.5

1.25

0.625

0.313

0.156

0.078

0.039

0.0195

0.0098

0

Replicate 1

4.833E+04

2.670E+04

2.013E+04

1.019E+04

4.769E+03

2.687E+03

1.385E+03

6.670E+02

3.450E+02

1.630E+02

6.800E+01

2.000E+00

Replicate 2

4.477E+04

3.233E+04

1.903E+04

1.032E+04

4.521E+03

2.770E+03

1.147E+03

5.660E+02

2.560E+02

1.950E+02

1.200E+02

4.000E+00

Replicate 3

4.932E+04

3.348E+04

2.032E+04

1.298E+04

6.118E+03

2.550E+03

1.468E+03

5.660E+02

2.710E+02

1.730E+02

7.900E+01

5.000E+00

Replicate 4

4.846E+04

3.634E+04

2.116E+04

1.180E+04

4.451E+03

3.568E+03

1.445E+03

7.330E+02

3.010E+02

1.310E+02

6.800E+01

8.000E+00

Arith. Mean

4.772E+04

3.221E+04

2.016E+04

1.132E+04

4.965E+03

2.894E+03

1.361E+03

6.330E+02

2.933E+02

1.655E+02

8.375E+01

4.750E+00

Arith. Std.Dev.

2.012E+03

4.042E+03

8.766E+02

1.323E+03

7.808E+02

4.586E+02

1.471E+02

8.192E+01

3.925E+01

2.660E+01

2.472E+01

2.500E+00

Arith. Net Mean

4.771E+04

3.221E+04

2.016E+04

1.132E+04

4.960E+03

2.889E+03

1.357E+03

6.283E+02

2.885E+02

1.608E+02

7.900E+01

Figure 3 : The fluorescein standard curve in log scale (excitation light:485nm, emission light 530nm)

Figure 4 : The fluorescein standard curve in line scale (excitation light:485nm, emission light 530nm)

The standard curve are straight line in both the line and log scale. So the standard curve may be credible after we use it in following experiment and the analyzing results.

Cell measurement

The fourth segment is the results of absorbance 600nm and fluorescence of transformed cells. According to the protocol we get the data and comparing them with the results in CFU/ml to get our conclusion whether CFU/ml could replace the absorbance in fluorescence/od in laboratory works.

Table 4: The raw data we get from the plate reader after measuring the samples from 0h and 6h during cultivation of the transformed cells.

Fluorescence Raw Readings:
Hour 0:	Neg. Control	Pos. Control	Device 1	Device 2	Device 3	Device 4	Device 5	Device 6	LB + Chlor (blank)
Colony 1, Replicate 1	126	108	128	197	180	293	169	119	64
Colony 1, Replicate 2	93	104	72	250	136	264	151	164	66
Colony 1, Replicate 3	100	112	123	233	122	276	135	157	70
Colony 1, Replicate 4	98	129	135	189	78	258	98	148	68
Colony 2, Replicate 1	87	130	138	176	161	223	134	163	79
Colony 2, Replicate 2	112	131	125	156	244	161	142	159	66
Colony 2, Replicate 3	103	125	89	132	159	255	160	158	64
Colony 2, Replicate 4	89	139	105	143	134	245	148	163	66

Hour 6:	Neg. Control	Pos. Control	Device 1	Device 2	Device 3	Device 4	Device 5	Device 6	LB + Chlor (blank)
Colony 1, Replicate 1	10203	18662	17149	18644	10574	22936	20801	13347	7858
Colony 1, Replicate 2	11003	19975	18619	19508	11235	23844	19238	13822	8292
Colony 1, Replicate 3	11132	20487	18620	19861	11685	24304	19073	14437	8165
Colony 1, Replicate 4	11888	21333	18801	20514	11712	29351	19624	14765	8494
Colony 2, Replicate 1	11077	17324	20389	18729	10855	25237	20372	15074	8274
Colony 2, Replicate 2	10855	18456	20509	18580	11291	25137	20489	15131	8684
Colony 2, Replicate 3	10900	18443	21305	19037	11065	24776	20467	15603	8563
Colony 2, Replicate 4	10386	18345	24383	18403	10966	25233	19799	15265	8544

Abs600 Raw Readings:
Hour 0:	Neg. Control	Pos. Control	Device 1	Device 2	Device 3	Device 4	Device 5	Device 6	LB + Chlor (blank)
Colony 1, Replicate 1	0.05	0.053	0.049	0.05	0.062	0.048	0.045	0.054	0.041
Colony 1, Replicate 2	0.049	0.05	0.048	0.052	0.052	0.046	0.044	0.048	0.04
Colony 1, Replicate 3	0.049	0.055	0.051	0.055	0.058	0.046	0.046	0.053	0.042
Colony 1, Replicate 4	0.043	0.047	0.052	0.049	0.054	0.049	0.045	0.051	0.044
Colony 2, Replicate 1	0.048	0.052	0.056	0.052	0.051	0.047	0.043	0.049	0.041
Colony 2, Replicate 2	0.054	0.058	0.052	0.049	0.05	0.051	0.046	0.052	0.043
Colony 2, Replicate 3	0.052	0.049	0.054	0.051	0.053	0.049	0.048	0.05	0.042
Colony 2, Replicate 4	0.051	0.047	0.047	0.051	0.051	0.048	0.05	0.054	0.041

Hour 6:	Neg. Control	Pos. Control	Device 1	Device 2	Device 3	Device 4	Device 5	Device 6	LB + Chlor (blank)
Colony 1, Replicate 1	0.313	0.224	0.22	0.239	0.271	0.229	0.214	0.24	0.043
Colony 1, Replicate 2	0.328	0.226	0.221	0.254	0.266	0.228	0.218	0.244	0.049
Colony 1, Replicate 3	0.294	0.218	0.217	0.25	0.264	0.234	0.216	0.252	0.043
Colony 1, Replicate 4	0.276	0.227	0.223	0.233	0.275	0.225	0.216	0.25	0.042
Colony 2, Replicate 1	0.279	0.236	0.238	0.264	0.249	0.255	0.206	0.205	0.044
Colony 2, Replicate 2	0.266	0.237	0.228	0.277	0.248	0.255	0.208	0.21	0.041
Colony 2, Replicate 3	0.27	0.236	0.236	0.264	0.241	0.252	0.202	0.202	0.05
Colony 2, Replicate 4	0.28	0.25	0.23	0.266	0.248	0.256	0.203	0.203	0.043

After synthesis the raw data that we get from the calibration protocol and cell measurement protocol, there are also some forms after analyzing. (for easily to compare the difference between the devices, there are some diagrams only present after cultivating 6h, because we think some data getting from samples in 0h is less valuable )

Table 5: The unit scaling factors of our standard curve in calibration experiment.

Unit Scaling Factors:
OD600 / Abs600	3.55
uM Fluorescein / a.u.	1.25E-04

Table 6: The net absorbance 600nm

	Net Abs 600
Hour 0:	Neg. Control	Pos. Control	Device 1	Device 2	Device 3	Device 4	Device 5	Device 6
Colony 1, Replicate 1	0.009	0.012	0.008	0.009	0.021	0.007	0.004	0.013
Colony 1, Replicate 2	0.009	0.010	0.008	0.012	0.012	0.006	0.004	0.008
Colony 1, Replicate 3	0.007	0.013	0.009	0.013	0.016	0.004	0.004	0.011
Colony 1, Replicate 4	-0.001	0.003	0.008	0.005	0.010	0.005	0.001	0.007
Colony 2, Replicate 1	0.007	0.011	0.015	0.011	0.010	0.006	0.002	0.008
Colony 2, Replicate 2	0.011	0.015	0.009	0.006	0.007	0.008	0.003	0.009
Colony 2, Replicate 3	0.010	0.007	0.012	0.009	0.011	0.007	0.006	0.008
Colony 2, Replicate 4	0.010	0.006	0.006	0.010	0.010	0.007	0.009	0.013

Hour 6:	Neg. Control	Pos. Control	Device 1	Device 2	Device 3	Device 4	Device 5	Device 6
Colony 1, Replicate 1	0.270	0.181	0.177	0.196	0.228	0.186	0.171	0.197
Colony 1, Replicate 2	0.279	0.177	0.172	0.205	0.217	0.179	0.169	0.195
Colony 1, Replicate 3	0.251	0.175	0.174	0.207	0.221	0.191	0.173	0.209
Colony 1, Replicate 4	0.234	0.185	0.181	0.191	0.233	0.183	0.174	0.208
Colony 2, Replicate 1	0.235	0.192	0.194	0.220	0.205	0.211	0.162	0.161
Colony 2, Replicate 2	0.225	0.196	0.187	0.236	0.207	0.214	0.167	0.169
Colony 2, Replicate 3	0.220	0.186	0.186	0.214	0.191	0.202	0.152	0.152
Colony 2, Replicate 4	0.237	0.207	0.187	0.223	0.205	0.213	0.160	0.160

Figure 5 : The net absorbance in 600nm of transformed cells after 6h cultivation

Table 7: The net Fluorescence data

Net Fluorescein a.u.
Hour 0:	Neg. Control	Pos. Control	Device 1	Device 2	Device 3	Device 4	Device 5	Device 6
Colony 1, Replicate 1	62.00	44.00	64.00	133.00	116.00	229.00	105.00	55.00
Colony 1, Replicate 2	27.00	38.00	6.00	184.00	70.00	198.00	85.00	98.00
Colony 1, Replicate 3	30.00	42.00	53.00	163.00	52.00	206.00	65.00	87.00
Colony 1, Replicate 4	30.00	61.00	67.00	121.00	10.00	190.00	30.00	80.00
Colony 2, Replicate 1	8.00	51.00	59.00	97.00	82.00	144.00	55.00	84.00
Colony 2, Replicate 2	46.00	65.00	59.00	90.00	178.00	95.00	76.00	93.00
Colony 2, Replicate 3	39.00	61.00	25.00	68.00	95.00	191.00	96.00	94.00
Colony 2, Replicate 4	23.00	73.00	39.00	77.00	68.00	179.00	82.00	97.00

Hour 6:	Neg. Control	Pos. Control	Device 1	Device 2	Device 3	Device 4	Device 5	Device 6
Colony 1, Replicate 1	2345.00	10804.00	9291.00	10786.00	2716.00	15078.00	12943.00	5489.00
Colony 1, Replicate 2	2711.00	11683.00	10327.00	11216.00	2943.00	15552.00	10946.00	5530.00
Colony 1, Replicate 3	2967.00	12322.00	10455.00	11696.00	3520.00	16139.00	10908.00	6272.00
Colony 1, Replicate 4	3394.00	12839.00	10307.00	12020.00	3218.00	20857.00	11130.00	6271.00
Colony 2, Replicate 1	2803.00	9050.00	12115.00	10455.00	2581.00	16963.00	12098.00	6800.00
Colony 2, Replicate 2	2171.00	9772.00	11825.00	9896.00	2607.00	16453.00	11805.00	6447.00
Colony 2, Replicate 3	2337.00	9880.00	12742.00	10474.00	2502.00	16213.00	11904.00	7040.00
Colony 2, Replicate 4	1842.00	9801.00	15839.00	9859.00	2422.00	16689.00	11255.00	6721.00

Figure 6 : The net fluorescein of transformed cells after 6h cultivation (excitation light:485nm, emission light 530nm)

Table 8: The um fluorescence/OD data

uM Fluorescein / OD
Hour 0:	Neg. Control	Pos. Control	Device 1	Device 2	Device 3	Device 4	Device 5	Device 6
Colony 1, Replicate 1	0.242	0.129	0.281	0.520	0.194	1.150	0.923	0.149
Colony 1, Replicate 2	0.105	0.134	0.026	0.539	0.205	1.160	0.747	0.431
Colony 1, Replicate 3	0.151	0.114	0.207	0.441	0.114	1.811	0.571	0.278
Colony 1, Replicate 4	-1.055	0.715	0.294	0.851	0.035	1.336	1.055	0.402
Colony 2, Replicate 1	0.040	0.163	0.138	0.310	0.288	0.844	0.967	0.369
Colony 2, Replicate 2	0.147	0.152	0.230	0.527	0.894	0.417	0.891	0.363
Colony 2, Replicate 3	0.137	0.306	0.073	0.266	0.304	0.959	0.562	0.413
Colony 2, Replicate 4	0.081	0.428	0.229	0.271	0.239	0.899	0.320	0.262


Figure 7 : The ratio of fluorescein/OD in 0h (excitation light:485nm, emission light 530nm)	Figure 8 : The ratio of fluorescein/OD after cultivation for 6h (excitation light:485nm, emission light 530nm)

Discussion: According to the net fluorescence per od value, the order of the strengths of the different promoters is estimated based on the average of two colonies in each group. The conclusion is that "device 4">"device 5">"device1" approximately equal to "positive control" >"device 2">"device 6">"device 3">"negative control". The results of the comparison with the official RFP were basically the same, while the positive and negative controls and the repetition of the two colonies indicated that the results were credible.

Table 9 : The unit scaling factors of our standard curve in calibration experiment.

Unit Scaling Factors:

Hour 6:	Neg. Control	Pos. Control	Device 1	Device 2	Device 3	Device 4	Device 5	Device 6
Colony 1, Replicate 1	0.305	2.098	1.845	1.935	0.419	2.850	2.661	0.980
Colony 1, Replicate 2	0.342	2.320	2.111	1.923	0.477	3.054	2.277	0.997
Colony 1, Replicate 3	0.416	2.475	2.112	1.986	0.560	2.971	2.217	1.055
Colony 1, Replicate 4	0.510	2.440	2.002	2.212	0.486	4.007	2.249	1.060
Colony 2, Replicate 1	0.419	1.657	2.195	1.671	0.443	2.826	2.625	1.485
Colony 2, Replicate 2	0.339	1.753	2.223	1.474	0.443	2.703	2.485	1.341
Colony 2, Replicate 3	0.373	1.867	2.408	1.721	0.461	2.822	2.753	1.628
Colony 2, Replicate 4	0.273	1.665	2.978	1.554	0.415	2.755	2.473	1.477

Particles / Abs600

4.00E+08

MEFL / a.u.

7.51E+08

Table 10: MEFL/particle

MEFL / particle
Hour 0:	Neg. Control	Pos. Control	Device 1	Device 2	Device 3	Device 4	Device 5	Device 6
Colony 1, Replicate 1	1.29E+04	6.89E+03	1.50E+04	2.78E+04	1.04E+04	6.15E+04	4.93E+04	7.95E+03
Colony 1, Replicate 2	5.64E+03	7.14E+03	1.41E+03	2.88E+04	1.10E+04	6.20E+04	3.99E+04	2.30E+04
Colony 1, Replicate 3	8.06E+03	6.07E+03	1.11E+04	2.36E+04	6.11E+03	9.68E+04	3.05E+04	1.49E+04
Colony 1, Replicate 4	-5.64E+04	3.82E+04	1.57E+04	4.55E+04	1.88E+03	7.14E+04	5.64E+04	2.15E+04
Colony 2, Replicate 1	2.15E+03	8.72E+03	7.39E+03	1.66E+04	1.54E+04	4.51E+04	5.17E+04	1.97E+04
Colony 2, Replicate 2	7.86E+03	8.15E+03	1.23E+04	2.82E+04	4.78E+04	2.23E+04	4.76E+04	1.94E+04
Colony 2, Replicate 3	7.33E+03	1.64E+04	3.92E+03	1.42E+04	1.62E+04	5.13E+04	3.01E+04	2.21E+04
Colony 2, Replicate 4	4.32E+03	2.29E+04	1.22E+04	1.45E+04	1.28E+04	4.81E+04	1.71E+04	1.40E+04

Hour 6:	Neg. Control	Pos. Control	Device 1	Device 2	Device 3	Device 4	Device 5	Device 6
Colony 1, Replicate 1	1.63E+04	1.12E+05	9.87E+04	1.03E+05	2.24E+04	1.52E+05	1.42E+05	5.24E+04
Colony 1, Replicate 2	1.83E+04	1.24E+05	1.13E+05	1.03E+05	2.55E+04	1.63E+05	1.22E+05	5.33E+04
Colony 1, Replicate 3	2.22E+04	1.32E+05	1.13E+05	1.06E+05	2.99E+04	1.59E+05	1.19E+05	5.64E+04
Colony 1, Replicate 4	2.73E+04	1.30E+05	1.07E+05	1.18E+05	2.60E+04	2.14E+05	1.20E+05	5.67E+04
Colony 2, Replicate 1	2.24E+04	8.86E+04	1.17E+05	8.93E+04	2.37E+04	1.51E+05	1.40E+05	7.94E+04
Colony 2, Replicate 2	1.81E+04	9.37E+04	1.19E+05	7.88E+04	2.37E+04	1.45E+05	1.33E+05	7.17E+04
Colony 2, Replicate 3	2.00E+04	9.99E+04	1.29E+05	9.20E+04	2.46E+04	1.51E+05	1.47E+05	8.71E+04
Colony 2, Replicate 4	1.46E+04	8.90E+04	1.59E+05	8.31E+04	2.22E+04	1.47E+05	1.32E+05	7.90E+04


Figure 9 : The ratio of fluorescein/particle in 0h (excitation light:485nm, emission light 530nm)	Figure 10 : The ratio of fluorescein/particle after cultivation for 6h (excitation light:485nm, emission light 530nm)

Discussion : MEFL/particle also show the same results that device 4 have the stongest promotor and device 3 have the weakest promotor. And the decreasing promotor strength is "device 4">"device 5">"device1" approximately equal to "positive control" >"device 2">"device 6">"device 3">"negative control", which corresopnd to the results in flourescence per od .

CFU/ml data in Neg.Control and Pos.Control .

We dilute the neg.control and pos.control into the absorbance into 0.1, and we finish the following dilution according to the protocol. The protocol is easy to understand and there presents the data we record during experiment.

Table 11: The original absorbance of neg.control and pos.control after dilution in plate reader

Neg.Control Relplicate 1	Neg.Control Relplicate 2	Neg.Control Relplicate 3	Pos.Control Relplicate 1	Pos.Control Relplicate 2	Pos.Control Relplicate 3	LB+Cam
0.148	0.154	0.153	0.142	0.158	0.145	0.042
0.157	0.165	0.159	0.143	0.135	0.139	0.041

Table 12: The colony numbers in plates

	Colony numbers
Final Dilution Factor	Neg.Control Relplicate 1	Neg.Control Relplicate 2	Neg.Control Relplicate 3	Pos.Control Relplicate 1	Pos.Control Relplicate 2	Pos.Control Relplicate 3
8*10^4	542	2300	1992	285	404	261
8*10^5	174	211	143	25	40	46
8*10^6	33	9	35	5	3	1
Final Dilution Factor	Neg.Control Relplicate 1	Neg.Control Relplicate 2	Neg.Control Relplicate 3	Pos.Control Relplicate 1	Pos.Control Relplicate 2	Pos.Control Relplicate 3
8*10^4	992	1488	660	267	209	221
8*10^5	140	288	79	16	22	22
8*10^6	8	53	16	2	13	1

Final CFU/ml	Neg.Control Relplicate 1	Neg.Control Relplicate 2	Neg.Control Relplicate 3	Pos.Control Relplicate 1	Pos.Control Relplicate 2	Pos.Control Relplicate 3
colony 1	2.01E+08	1.69E+08	1.97E+08	2.28E+07	3.20E+07	2.88E+07
colony 2	1.12E+08	3.27E+08	3.92E+07	2.14E+07	1.67E+07	1.77E+07

Figure 11 : The colony numbers per milliliter in 0.1 absorbance culture (600nm)

we selectively choose some data to calculate the final CFU/ml . The selective standard is according to the plate colony numbers which is between 30-300. After analyzing the final CFU/ml in the negative and positive control, we found that although the origin absorance are similar, there are some difference between the CFU/ml. we are not sure whether there exists pipetting errors during dilution process . But all the data we got shows that the expressing GFP consistantly in Pos.Control may affect the growth of bacteria .

Final conclusion

We could conclude that there are some relationships between the fluorescence/OD and fluorescence per particle, So it is definitely that we could use particle of cells to replace the Absorbance of cells. However, there is no obvious relationship between the particles of cells and the CFU/ml in culture. Because CFU/ml presents the numbers of survival cells and the bacteria which have relatively strong vitality. Compared with positive control, negative control obviously have a higher chance to express antibiotic resistant gene instead of the unnecessary GFP. So when transformed cells grow on the plates, there are more chances to survival in the antibiotic plates for cells who express more less unnecessary proteins.

Of course, there may exists some pipetting errors during dilution process, but the data we obtain from the experiment show that we may can’t normalize colony-forming units (CFUs) instead of OD during fluorescence measurements .


Figure 8 : The ratio of fluorescein/OD after cultivation for 6h (excitation light:485nm, emission light 530nm)	Figure 10 : The ratio of fluorescein/particle after cultivation for 6h (excitation light:485nm, emission light 530nm)

Figure 11 : The colony numbers per milliliter in 0.1 absorbance culture (600nm)

@@ Line 439: / Line 439: @@
 <body lang=ZH-CN style='tab-interval:21.0pt;text-justify-trim:punctuation'>
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+<div class=WordSection1 style='layout-grid: 15.6pt; text-align: left;'>
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+<div>
+ <img src="https://static.igem.org/mediawiki/2018/f/fa/T--WHU-China--interlab-image001.png" width="866" height="454">
+  <p style='color: red;'>Figure 1：The Particle Standard  Curve with monodisperse silica microspheres (log scale) </p>
+</div>
+  <div>
+ <img src="https://static.igem.org/mediawiki/2018/f/f6/T--WHU-China--interlab-image003.png" width="866" height="454">
+  <p style='color: red;'>Figure 2 : The Particle Standard Curve with monodisperse silica microspheres (line scale) </p>
+</div>
 <p class=MsoNoSpacing><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
 <p class="Table-title"style='color:red'>Table 3: The raw data  after measuring the fluorescence (excitation light: 485 nm, emission light  530nm) in plate reader.</p>
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 </div>
 <p class=MsoNoSpacing><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
-<p class=MsoNormal><span lang=EN-US style='mso-no-proof:yes'><![endif]></span><span class="GramE"><span style="mso-no-proof:yes"><img width=554 height=283
+<div>
-src="https://static.igem.org/mediawiki/2018/1/1b/T--WHU-China--interlab-image008.png" v:shapes="图表_x0020_4"></span></span></p>
+ <img src="https://static.igem.org/mediawiki/2018/d/d5/T--WHU-China--interlab-image007.png" width="866" height="454">
+  <p style='color: red; '>Figure 3 : The fluorescein standard curve in log scale (excitation light:485nm, emission light 530nm)</p>
+</div>
+<div>
+ <img src="https://static.igem.org/mediawiki/2018/9/99/T--WHU-China--interlab-image005.png" width="866" height="454">
+  <p style='color: red; '>Figure 4 : The fluorescein standard curve in line scale (excitation light:485nm, emission light 530nm) </p>
+</div>
 <p class=MsoNormal><span lang=EN-US><o:p>&nbsp;</o:p></span></p>
@@ Line 1,564: / Line 1,578: @@
 </div></p>
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+<div>
-lang=EN-US style='font-size:12.0pt;font-family:宋体;mso-bidi-font-family:宋体;
+  <div ><img src="https://static.igem.org/mediawiki/2018/0/0b/T--WHU-China--interlab-image009.png" width="520" height="312">
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+ </div>
- o:spid="_x0000_i1034" type="#_x0000_t75" style='width:414.6pt;height:249pt;
+ <p style='color: red; '>Figure 5 : The net absorbance in 600nm of transformed cells after 6h cultivation  </p>
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+</div>
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 <p class="Table-title"style='color:red'>Table 7: The net  Fluorescence data </p>
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      <td valign="bottom" nowrap="nowrap" bgcolor="#FF9900"><p align="right">97.00</p></td>
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 </div>
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 mso-font-kerning:0pt'><o:p>&nbsp;</o:p></span></p>
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+  <div><img src="https://static.igem.org/mediawiki/2018/f/f2/T--WHU-China--interlab-image011.png" width="520" height="312">
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+ </div>
- o:spid="_x0000_i1033" type="#_x0000_t75" style='width:414.6pt;height:249pt;
+ <p style='color: red;'>Figure 6 : The net fluorescein of transformed cells after 6h cultivation (excitation light:485nm, emission light 530nm) </p>
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+<div>
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+    <table width="100%" border="0.1">
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+      <tr>
- <v:imagedata src="interlab.files/image013.png" o:title="7EJ9PMUIEU_J%_52(JG_LJX"/>
+        <td width="61%"><img src="https://static.igem.org/mediawiki/2018/3/35/T--WHU-China--interlab-image013.png" width="481" height="289"></td>
-</v:shape><![endif]--><![if !vml]><img width=553 height=332
+        <td width="39%"><img src="https://static.igem.org/mediawiki/2018/7/7b/T--WHU-China--interlab-image015.png" width="481" height="289"></td>
-src="https://static.igem.org/mediawiki/2018/a/a5/T--WHU-China--interlab-image014.jpg" v:shapes="图片_x0020_5"></span></p>
+      </tr>
-<p class=MsoNormal>&nbsp;</p>
+      <tr>
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+        <td><div align="center">
-mso-bidi-font-family:宋体;mso-font-kerning:0pt;mso-no-proof:yes'><![endif]></span><span
+          <p style="color: red;">Figure 7 : The ratio of fluorescein/OD in 0h (excitation light:485nm, emission light 530nm) </p>
-lang=EN-US style='mso-no-proof:yes'><!--[if gte vml 1]><v:shape id="图片_x0020_6"
+        </div></td>
- o:spid="_x0000_i1031" type="#_x0000_t75" style='width:414.6pt;height:249pt;
+        <td><div align="center"><span style="color: red;">Figure 8 : The ratio of fluorescein/OD after cultivation for 6h (excitation light:485nm, emission light 530nm) </span></div></td>
- visibility:visible;mso-wrap-style:square'>
+      </tr>
- <v:imagedata src="interlab.files/image015.png" o:title="ZOR[M8%J8Y@S`~W_QGRDPQU"/>
+    </table>
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+  </div>
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 </div>
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+      <tr>
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+        <td width="61%"><img src="https://static.igem.org/mediawiki/2018/9/9d/T--WHU-China--interlab-image017.png" width="481" height="289"></td>
+        <td width="39%"><img src="https://static.igem.org/mediawiki/2018/9/91/T--WHU-China--interlab-image019.png" width="481" height="289"></td>
-<p class=MsoNormal><span lang=EN-US style='mso-no-proof:yes'><![if !vml]></span></p>
+      </tr>
-<p class=MsoNormal><span lang=EN-US style='mso-no-proof:yes'><img width=554 height=331
+      <tr>
-src="https://static.igem.org/mediawiki/2018/a/a2/T--WHU-China--interlab-image020.png" v:shapes="图表_x0020_13"><![endif]></span><span
+        <td><div align="center">
-lang=EN-US style='color:red'><o:p></o:p></span></p>
+          <p style="color: red;">Figure 9 : The ratio of fluorescein/particle in 0h (excitation light:485nm, emission light 530nm) </p>
+        </div></td>
+        <td><div align="center"><span style="color: red;">Figure 10 : The ratio of fluorescein/particle after cultivation for 6h (excitation light:485nm, emission light 530nm) </span></div></td>
+      </tr>
+    </table>
+  </div>
+</div>
 <p class=MsoNormal><span lang=EN-US style='color:red'><o:p>&nbsp;</o:p></span></p>
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   <v:imagedata src="interlab.files/image021.png" o:title=""/>
 </v:shape><![endif]--><![if !vml]></span></span></p>
-<p class=MsoNormal><span lang=EN-US><span style='mso-no-proof:yes'><img width=554 height=333
+<div>
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+  </div>
-  DrawAspect="Content" ObjectID="_1593884745">
+  <p style='color: red; text-align: center;'>Figure 11 : The colony numbers per milliliter in 0.1 absorbance culture (600nm)  </p>
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 <span class=GramE>measurements .</span></span></p>
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+<div>
-style='mso-no-proof:yes'><![if !vml]><img width=553 height=351
+  <div align="left">
-src="https://static.igem.org/mediawiki/2018/c/c8/T--WHU-China--interlab-image024.png" v:shapes="图表_x0020_17"><![endif]></span><span style='mso-no-proof:yes'><![if !vml]><img width=553 height=351
+    <table border="0.1">
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+      <tr>
-src="https://static.igem.org/mediawiki/2018/5/58/T--WHU-China--interlab-image027.png" v:shapes="图表_x0020_16"><![endif]></span></span></p>
+        <td><img src="https://static.igem.org/mediawiki/2018/7/7b/T--WHU-China--interlab-image015.png" width="481" height="289"></td>
+        <td><img src="https://static.igem.org/mediawiki/2018/9/91/T--WHU-China--interlab-image019.png" width="481" height="290"></td>
+      </tr>
+      <tr>
+        <td><div align="center">
+          <p style="color: red;">Figure 8 : The ratio of fluorescein/OD after cultivation for 6h (excitation light:485nm, emission light 530nm) </p>
+        </div></td>
+        <td><div align="center"><span style="color: red;">Figure 10 : The ratio of fluorescein/particle after cultivation for 6h (excitation light:485nm, emission light 530nm) </span></div></td>
+      </tr>
+      <tr>
+        <td colspan="2"><img src="https://static.igem.org/mediawiki/2018/6/65/T--WHU-China--interlab-image021.png" width="966" height="582"></td>
+        </tr>
+      <tr>
+        <td colspan="2"><div><span style="color: red;">Figure 11 : The colony numbers per milliliter in 0.1 absorbance culture (600nm)  </span></div></td>
+        </tr>
+    </table>
+  </div>
+</div>
+</div>
 </div>

Difference between revisions of "Team:WHU-China/InterLab"