Difference between revisions of "Team:AHUT China/Demonstrate"

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         <style>
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cursor: crosshair;
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display: block;
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}
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</style>
 
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         <!--================Header Area =================-->
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         <section class="breadcrumb_area">
 
         <section class="breadcrumb_area">
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       <script src="https://2018.igem.org/Team:AHUT_China/contact_js?action=raw&ctype=text/javascript" type="text/javascript" ></script>
 
       <script src="https://2018.igem.org/Team:AHUT_China/contact_js?action=raw&ctype=text/javascript" type="text/javascript" ></script>
 
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       <script src="https://2018.igem.org/Team:AHUT_China/custom_js?action=raw&ctype=text/javascript" type="text/javascript" ></script>
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 +
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 +
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 +
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window.mozRequestAnimationFrame ||
 +
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 +
window.setTimeout( callback, 1000 / 60 );
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};
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})();
 +
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// now we will setup our basic variables for the demo
 +
var canvas = document.getElementById( 'canvas' ),
 +
ctx = canvas.getContext( '2d' ),
 +
// full screen dimensions
 +
cw = window.innerWidth,
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 +
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 +
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// now we are going to setup our function placeholders for the entire demo
 +
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 +
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 +
return Math.random() * ( max - min ) + min;
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 +
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 +
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 +
var xDistance = p1x - p2x,
 +
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 +
return Math.sqrt( Math.pow( xDistance, 2 ) + Math.pow( yDistance, 2 ) );
 +
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 +
 +
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 +
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 +
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 +
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 +
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Firework.prototype.update = function( index ) {
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 +
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 +
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 +
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 +
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// remove the firework, use the index passed into the update function to determine which to remove
 +
fireworks.splice( index, 1 );
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 +
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 +
this.x += vx;
 +
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 +
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 +
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 +
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 +
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function Particle( x, y ) {
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this.x = x;
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this.y = y;
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 +
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 +
this.coordinates.push( [ this.x, this.y ] );
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 +
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 +
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 +
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 +
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 +
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 +
 +
 +
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 +
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 +
 
 +
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 +
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 +
ctx.moveTo( this.coordinates[ this.coordinates.length - 1 ][ 0 ], this.coordinates[ this.coordinates.length - 1 ][ 1 ] );
 +
ctx.lineTo( this.x, this.y );
 +
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 +
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 +
 
 +
 
 +
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 +
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 +
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 +
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 +
var particleCount = 30;
 +
while( particleCount-- ) {
 +
particles.push( new Particle( x, y ) );
 +
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 +
}
 +
 +
// main demo loop
 +
function loop() {
 +
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 +
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 +
 +
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 +
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 +
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// normally, clearRect() would be used to clear the canvas
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// we want to create a trailing effect though
 +
// setting the composite operation to destination-out will allow us to clear the canvas at a specific opacity, rather than wiping it entirely
 +
ctx.globalCompositeOperation = 'destination-out';
 +
// decrease the alpha property to create more prominent trails
 +
ctx.fillStyle = 'rgba(0, 0, 0, 0.5)';
 +
ctx.fillRect( 0, 0, cw, ch );
 +
// change the composite operation back to our main mode
 +
// lighter creates bright highlight points as the fireworks and particles overlap each other
 +
ctx.globalCompositeOperation = 'lighter';
 +
 +
    var text = "Welcom to our pages"; 
 +
  ctx.font = "50px sans-serif";
 +
  var textData = ctx.measureText(text);
 +
  ctx.fillStyle = "rgba("+parseInt(random(0,255))+","+parseInt(random(0,255))+","+parseInt(random(0,255))+",0.3)";
 +
  ctx.fillText(text,cw /2-textData.width/2,ch/2);
 +
 
 +
// loop over each firework, draw it, update it
 +
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 +
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 +
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 +
fireworks[ i ].update( i );
 +
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 +
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// loop over each particle, draw it, update it
 +
var i = particles.length;
 +
while( i-- ) {
 +
particles[ i ].draw();
 +
particles[ i ].update( i );
 +
}
 +
 +
// launch fireworks automatically to random coordinates, when the mouse isn't down
 +
if( timerTick >= timerTotal ) {
 +
if( !mousedown ) {
 +
// start the firework at the bottom middle of the screen, then set the random target coordinates, the random y coordinates will be set within the range of the top half of the screen
 +
     
 +
      for(var h=0;h<50;h++)
 +
      {
 +
          fireworks.push( new Firework( cw / 2, ch/2, random( 0, cw ), random( 0, ch  ) ) );
 +
      }
 +
     
 +
     
 +
timerTick = 0;
 +
}
 +
} else {
 +
timerTick++;
 +
}
 +
 +
// limit the rate at which fireworks get launched when mouse is down
 +
if( limiterTick >= limiterTotal ) {
 +
if( mousedown ) {
 +
// start the firework at the bottom middle of the screen, then set the current mouse coordinates as the target
 +
fireworks.push( new Firework( cw / 2, ch/2, mx, my ) );
 +
limiterTick = 0;
 +
}
 +
} else {
 +
limiterTick++;
 +
}
 +
}
 +
 +
// mouse event bindings
 +
// update the mouse coordinates on mousemove
 +
canvas.addEventListener( 'mousemove', function( e ) {
 +
mx = e.pageX - canvas.offsetLeft;
 +
my = e.pageY - canvas.offsetTop;
 +
});
 +
 +
// toggle mousedown state and prevent canvas from being selected
 +
canvas.addEventListener( 'mousedown', function( e ) {
 +
e.preventDefault();
 +
mousedown = true;
 +
});
 +
 +
canvas.addEventListener( 'mouseup', function( e ) {
 +
e.preventDefault();
 +
mousedown = false;
 +
});
 +
 +
// once the window loads, we are ready for some fireworks!
 +
window.onload = loop;
 +
 +
</script>
 
</html>
 
</html>

Revision as of 03:35, 6 October 2018

Royal Hotel Royal Hotel

Demonstrate

  1. Home
  2. Demonstrate

     Introduction

We took part in the Fifth International InterLab Measurement Study which ains to achieve the purpose of comparative measurement. The goal of this study is to obtain large amounts of data from labs across the world,to develop absolute units for measurements GFP in a plate reader to eliminate variation between labs.


     Materials

Plate reader: Synergy H1 (Biotek)
Plate reader plates: Corning 3603 96-Well Microplates (black plates with clear flat bottom)
Cell culture shaker: ZWYR-200D

Devices:
Negative control ：BBa_R0040
Positive control ：BBa_I20270
Device 1： BBa_J364000
Device 2： BBa_J364001
Device 3： BBa_J364002
Device 4： BBa_J364007
Device 5： BBa_J364008
Device 6： BBa_J364009
Note: for Device 5, we have not transformed it into DH5⍺ competent cells successfully for many times, therefore, we thank IGEM team of Nanjing University for providing the Device 5.
Calibration material: Provided in the 2018 IGEM distribution kit
Microorganism: Escherichia coli DH5⍺ strains


     Methods

Following iGEM requirements, Team AHUT_China performed measurements according to these 2018 InterLab Protocols https://static.igem.org/mediawiki/2018/0/09/2018_InterLab_Plate_Reader_Protocol.pdf


     Results

1.OD 600 reference point

Using OD 600 and H2O to generate the conversion factor for the transformation later. The average of OD600 is 0.063; the correction factor (OD600/ABS600) is 3.500



Fig. 1 LUDOX correct value

2.Particle standard curve

We obtained the two Particle Standard Curve (normal and log scale).



Fig. 2 Particle Standard Curve

Fig. 3 Particle Standard Curve (log scale)

3.Fluorescein standard curve

Dilution serious of fluorescein were prepared and measured in a 96 well plate. A standard curve is generated to correct the cell based readings to an equivalent fluorescein concentration.
We obtained the two Fluorescein Standard Curve (normal and log scale).



Fig. 4 Fluorescein Standard Curve

Fig. 5 Fluorescein Standard Curve (log scale)

4.Cell measurements

 



Fig. 6 Fluorescence Measurements Curve

Test devices 1 and 4 show high fluorescence intensity. Test devices 2 show a modest fluorescence intensity alone with positive control group, while devices3,5,6 barely show low fluorescence intensity alone with the negative control group.



Fig. 7 Raw OD600 Curve

5.We obtained the Colony Forming Units per 0.1 OD600 E. coli cultures



Fig. 8 CFU Result

 

Discussion

For Figure 3, the log graph isn’t a straight line but not 1:1 slope. In figure 6, highest fluorescence was obtained from device 4, closely followed by test device 1. Test device 2 and positive control group show a modest fluorescence intensity and device 5,6 show low fluorescence intensity, while test devices 3 barely have any fluorescence signal as well as the negative group.

Conclusion

It was certainly a technical challenge to Participate in the InterLab Study. Performing the prescribed protocols with adherence to all the InterLab guidelines yielded parts of expected results, and with the completed InterLab Google Forms, confirms our team participation in this InterLab Study.