Team:SJTU-BioX-Shanghai/Protocol

• HOME
• PROJECT
  • Description
  • Design
  • Results
  • Demonstration
  • Loader
• EXPERIMENT
  • Parts
  • Notebook
  • Protocol
  • Improvement
  • Interlab
•  MODEL 
  • Overview
  • Adhesion
  • Ultrasonography
  • Gene Circuit
• PRACTICES
  • Background
  • Integrated
  • Engagement
  • Biosafety
  • Interview
  • Price Model
  • Application
•  TEAM 
  • Members
  • Attributions
  • Collaborations
• AWARDS

• Section1
• Section2
• Section3
• Section4
• Section5

Protocol

Section1

• Western Blotting protocol

  1、Prepare of Proteins
  1.1 Culture E.coli BL21(DE3) with LB(l) medium overnight at 37℃, 200rpm
  1.2 Add 1ml overnight culture into 5ml new LB(l) medium for 1 hour at 37℃, 200rpm
  1.3 Induce culture by IPTG (with final concentration of 0.5mM) more than 4 hours, while the OD600 is 0.6
  1.4 (Lysis) Add 1.5ml lysozyme (1mg/ml) into 2ml centrifuged and the supernatant discadred strain, 30℃ for 30 minutes
  1.5 (Degeneration) Add SDS loading buffer and put the centrifugal tube into boiled water for 10 minutes
  2、Prepare SDS-PAGE gel, Running buffer, Trans buffer and TBST Washing buffer
  SDS-PAGE:
  12% Separation Gel(the lower gel):
  30%Acr-Bis      3.2ml
  1.5M Tris-HCl(pH=8.8) 2ml
  10%SDS         80μl
  10%AP          80μl
  TEMED         3μl
  H2O         2.64ml
  Total         8ml
  5% Spacer Gel(the upper gel):
  30%Acr-Bis      500μl
  1.5M Tris-HCl(pH=6.8) 500μl
  10%SDS         40μl
  10%AP         30μl
  TEMED         3μl
  H2O         2ml
  Total         3ml
  1×Running buffer:
  Tris      3.03g
  Glycine    14.4g
  10%SDS    10ml
  Add ultra pure water to 1L
  Store at 4℃
  1×Trans buffer:
  Tris      3.03g
  Glycine    14.4g
  10%SDS     10ml
  Methanol    200ml
  Add ultra pure water to 1L
  Store at 4℃
  1×TBS buffer:
  Tris      2.4g
  NaCl     8.7g
  Adjust pH=7.6 with HCl
  Add ultra pure water to 1L
  TBST Washing buffer:
  TBS        500ml
  Tween-20     250μl
  Blocking buffer:
  Add 2.5g no-fat dry milk into 50ml TBST Washing buffer
  Prepare Blocking buffer only when using it
  3、Protein transfer and Immunodetection
  3.1 Add 15μl sample and 5μl Protein marker
  3.2 Add Running buffer for electrophoresis with a constant voltage at 100V for about 25 minutes, then change voltage into 200V for 40 minutes (adjust the former time by the marker)
  3.3 Open the package and cut the upper spacer gel
  3.4 Prepare the same size PVDF membrane and put into Trans buffer for 5 minutes before trans-membrane
  3.5 Open gel cassettes and put the gel on the PVDF membrane sandwiched between two pieces of blotting paper(always filter paper)
  Mind the order of gel and PVDF membrane between the electrodes, black end with gel for negative electrode and white end with PVDF membrane for positive electrode.
  3.6 Add Trans buffer for electrophoresis with a constant current at 350mA for 65 minutes(Crowd with ice at a 4℃ temperature )
  3.7 After trans-membrane, wash PVDF membrane with 10ml TBST for 10 minutes with gentle shaking
  3.8 Add 10ml Blocking buffer with gentle shaking for 1 hour at room temperature or 4℃ overnight
  3.9 Wash PVDF membrane with 10ml TBST for 10 minutes with gentle shaking
  3.10 Add 10ml diluted(1:5000) primary antibody for 2 hours at room temperature or 4℃ overnight with gentle shaking (primary antibody can be used repeatedly for 10 times)
  3.11 repeat step 3.9
  3.12 Add 10ml diluted secondary conjugated antibody for 2 hours at room temperature or 4℃ overnight with gentle shaking (seconfary antibody needn’t be repeatedly used)
  3.13 repeat step 3.9
  3.14 With the ratio of 1:1 to mix Luminous substrate A and B, typically 200-400μl mixture added on PVDF membrane
  3.15 Take photos by specific machine and store it
  

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 .

Mouse experiments

• CRC induction

  Subject: C57BL/6 mice strain (6~8 week old)
  Material: Azoxymethane (AOM) solution (1mg/ml)
  Dextran Sulfate Sodium Salt (DSS) solution (2.0% & 2.5%)
  Experiment aim:
  After analyzing the qualities of our device that requires testifying meanwhile thoroughly considering the 3Rs principle, we have reduced the need of animal experiments to the least, but for certain qualities the testifying procedure is difficult to replace, refine and reduce experimental animals use. As a result the colorectal cancer mouse model was induced in the minimum amount just to be adequate for product testifying procedure.
  Experiment design:
  Azoxymethane (AOM) is a chemical agent that can initiate cancer by alkylation of DNA, there by facilitating base mispairings, while Dextran Sulfate Sodium Salt (DSS) is a substance that causes inflammations that will eventually lead to enteritidis.
  After being injected into the subject, AOM introduces a carcinogenic environment in vivo, along with DSS consumed, enteritidis is likely to occur and soon develop into colorectal cancer.

• Your title2

  xxx xx xxxx xxx xxx xxx xx xxxx xxx xxxxx xxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxx

• Your title3

  xxx xx xxxx xxx xxx xxx xx xxxx xxx xxxxx xxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxx

Section3

xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx 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 text-link template is here.

2018 Interlab Plate Reader Protocol
Protocols/Transformation

• Western Blotting protocol

  1、Prepare of Proteins
  1.1 Culture E.coli BL21(DE3) with LB(l) medium overnight at 37℃, 200rpm
  1.2 Add 1ml overnight culture into 5ml new LB(l) medium for 1 hour at 37℃, 200rpm
  1.3 Induce culture by IPTG (with final concentration of 0.5mM) more than 4 hours, while the OD600 is 0.6
  1.4 (Lysis) Add 1.5ml lysozyme (1mg/ml) into 2ml centrifuged and the supernatant discadred strain, 30℃ for 30 minutes
  1.5 (Degeneration) Add SDS loading buffer and put the centrifugal tube into boiled water for 10 minutes
  2、Prepare SDS-PAGE gel, Running buffer, Trans buffer and TBST Washing buffer
  SDS-PAGE:
  12% Separation Gel(the lower gel):
  30%Acr-Bis      3.2ml
  1.5M Tris-HCl(pH=8.8) 2ml
  10%SDS         80μl
  10%AP          80μl
  TEMED         3μl
  H2O         2.64ml
  Total         8ml
  5% Spacer Gel(the upper gel):
  30%Acr-Bis      500μl
  1.5M Tris-HCl(pH=6.8) 500μl
  10%SDS         40μl
  10%AP         30μl
  TEMED         3μl
  H2O         2ml
  Total         3ml
  1×Running buffer:
  Tris      3.03g
  Glycine    14.4g
  10%SDS    10ml
  Add ultra pure water to 1L
  Store at 4℃
  1×Trans buffer:
  Tris      3.03g
  Glycine    14.4g
  10%SDS     10ml
  Methanol    200ml
  Add ultra pure water to 1L
  Store at 4℃
  1×TBS buffer:
  Tris      2.4g
  NaCl     8.7g
  Adjust pH=7.6 with HCl
  Add ultra pure water to 1L
  TBST Washing buffer:
  TBS        500ml
  Tween-20     250μl
  Blocking buffer:
  Add 2.5g no-fat dry milk into 50ml TBST Washing buffer
  Prepare Blocking buffer only when using it
  3、Protein transfer and Immunodetection
  3.1 Add 15μl sample and 5μl Protein marker
  3.2 Add Running buffer for electrophoresis with a constant voltage at 100V for about 25 minutes, then change voltage into 200V for 40 minutes (adjust the former time by the marker)
  3.3 Open the package and cut the upper spacer gel
  3.4 Prepare the same size PVDF membrane and put into Trans buffer for 5 minutes before trans-membrane
  3.5 Open gel cassettes and put the gel on the PVDF membrane sandwiched between two pieces of blotting paper(always filter paper)
  Mind the order of gel and PVDF membrane between the electrodes, black end with gel for negative electrode and white end with PVDF membrane for positive electrode.
  3.6 Add Trans buffer for electrophoresis with a constant current at 350mA for 65 minutes(Crowd with ice at a 4℃ temperature )
  3.7 After trans-membrane, wash PVDF membrane with 10ml TBST for 10 minutes with gentle shaking
  3.8 Add 10ml Blocking buffer with gentle shaking for 1 hour at room temperature or 4℃ overnight
  3.9 Wash PVDF membrane with 10ml TBST for 10 minutes with gentle shaking
  3.10 Add 10ml diluted(1:5000) primary antibody for 2 hours at room temperature or 4℃ overnight with gentle shaking (primary antibody can be used repeatedly for 10 times)
  3.11 repeat step 3.9
  3.12 Add 10ml diluted secondary conjugated antibody for 2 hours at room temperature or 4℃ overnight with gentle shaking (seconfary antibody needn’t be repeatedly used)
  3.13 repeat step 3.9
  3.14 With the ratio of 1:1 to mix Luminous substrate A and B, typically 200-400μl mixture added on PVDF membrane
  3.15 Take photos by specific machine and store it
  

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 .

Section

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

• Your title1

  xxx xx xxxx xxx xxx xxx xx xxxx xxx xxxxx xxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxx

• Your title2

  xxx xx xxxx xxx xxx xxx xx xxxx xxx xxxxx xxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxx

• Your title3

  xxx xx xxxx xxx xxx xxx xx xxxx xxx xxxxx xxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxxx xx xxxx xxx xxxxxxx

Section3

xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx xxx 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 text-link template is here.

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.

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

US imaging

We use the Fujifilm VisualSonics / VEVO LAZR-X imaging system and MX250 transducer. The application is set to VA Phantom. Parameters are listed below:

• Frequency: 21MHz
• Power for imaging: min(2%)
• Power for collapse: max(100%) (sustaining for 5 minutes)
• Max frame: 50
• Gain: 16dB
• Focus: adjust to the depth of the layer where engineered E.coli lies in.
• Field of view: adjust to achieve the maximum repression of noise and also zoom in on the interest region as much as possible.

Preparation before imaging: PBS solution is needed to cover the imaging samples allowing the transducer to dip into the solution. Pay attention to not producing any unwanted bubbles.

Copyright © 2018 SJTU-BioX-Shanghai. All rights reserved.