BME100 s2014:W Group9 L5: Difference between revisions
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'''Single-Drop Fluorimeter'''<br> | '''Single-Drop Fluorimeter'''<br> | ||
The Single-Drop Fluorimeter | The Single-Drop Fluorimeter is a small and black plastic box with two notches where the multi-welled slide can fit in. It has a switch located on the right hand side that allows for the blue LED light to be turned on or off. The slide of the fluorometer is aligned so that the blue LED shines through a drop placed over two middle wells. The slide has holes along it to allow for a sample to be placed in them. Placing the drops must be carefully done to ensure no overflow. | ||
<br> | <br> | ||
[[ | [[Image:Group9 Fluorometer.JPG]] | ||
<br> | <br> | ||
(This image was borrowed from another group because we did not take a picture of our set-up.) | |||
<br> | <br> | ||
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'''Smart Phone Camera Settings'''<br> | '''Smart Phone Camera Settings'''<br> | ||
* Type of Smartphone: iPhone 4s | * Type of Smartphone: iPhone 4s | ||
** Flash: Off | ** Flash: Off | ||
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'''Calibration'''<br> | '''Calibration'''<br> | ||
'' | '' For the calibration, the phone was placed into the cradle and the self-timer was readied. The flourimeter was 6 cm away from where the drop was going to be placed and the slide was placed into the holder. A sample of DNA was placed onto the slide using the micropipette. The black box was placed over the cradle and flourimeter and the picture was taken. This process was repeated for every DNA sample. | ||
* Distance between the smart phone cradle and drop = | * Distance between the smart phone cradle and drop = 6 cm | ||
'''Solutions Used for Calibration' | |||
'''Solutions Used for Calibration''' | |||
{| {{table}} width=700 | {| {{table}} width=700 | ||
|- | |- | ||
| | | 5 || 80 || 80 || 2.5 | ||
|- | |- | ||
| | | 2 || 80 || 80 || 1 | ||
|- | |- | ||
| | | 1 || 80 || 80 || 0.5 | ||
|- | |||
| 0.5 || 80 || 80 || 0.25 | |||
|- | |||
| 0.25 || 80 || 80 || 0.125 | |||
|- | |||
| 0 || 80 || 80 || 0 | |||
|} | |} | ||
<br> | |||
'''Placing Samples onto the Fluorimeter''' | '''Placing Samples onto the Fluorimeter''' | ||
# | # Insert the multi-welled slide into the notches on the fluorimeter so that the light passes through the middle of two rows. | ||
# | # Using the pipette, drop 80 μL of SYBR GREEN onto the slide. | ||
# | # Drop 80 μL of the DNA sample onto the same droplet, creating a mixture. | ||
# | # Cover the apparatus using the black box to prevent any light from creeping in and take a timed picture using a smartphone. | ||
<br> | <br> | ||
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'''Representative Images of Samples''' | '''Representative Images of Samples''' | ||
[[Image:Screen_Shot_2014-04-15_at_9.34.39_PM.png]] <br> | |||
The picture above is a sample with no DNA. <br> | |||
[[Image:Screen_Shot_2014-04-15_at_9.34.13_PM.png]] <br> | |||
The picture above is a sample with DNA. | |||
'''Image J Values for All Samples''' | '''Image J Values for All Samples''' | ||
<br> | |||
'' | {| | ||
| align="center" style="background:#f0f0f0;"|'''Final DNA concentration in SYBR Green I solution (µg/mL)''' | |||
| align="center" style="background:#f0f0f0;"|'''AREA''' | |||
| align="center" style="background:#f0f0f0;"|'''Mean Pixel Value''' | |||
| align="center" style="background:#f0f0f0;"|'''RAWINTDEN''' | |||
| align="center" style="background:#f0f0f0;"|'''RAWINTDEN''' | |||
|- | |||
| ||||||OF THE DROP||OF THE BACKGROUND | |||
|- | |||
| 1. 0||96384||7.169||690977||282517 | |||
|- | |||
| 2. 0||99668||6.198||617696||276082 | |||
|- | |||
| 3. 0||99532||7.54||750507||286709 | |||
|- | |||
| 4. 0.25||90952||7.788||708329||304315 | |||
|- | |||
| 5.0.25||77596||15.616||1211707||236426 | |||
|- | |||
| 6. 0.25||106672||4.01||427741||315565 | |||
|- | |||
| 7. 0.5||74072||10.234||758023||280597 | |||
|- | |||
| 8. 0.5||105444||20.949||2208918||318261 | |||
|- | |||
| 9. 0.5||115664||17.723||2049928||327516 | |||
|- | |||
| 10. 1||123672||38.948||4816732||460646 | |||
|- | |||
| 11. 1||121536||39.985||4859576||447692 | |||
|- | |||
| 12. 1||120572||42.031||5067789||457714 | |||
|- | |||
| 13. 2||123192||68.808||8476560||340345 | |||
|- | |||
| 14. 2||79768||149.736||11944117||269685 | |||
|- | |||
| 15. 2||121772||59.096||7196188||451155 | |||
|- | |||
| 16. 5 ||155476||52.53||8167202||593385 | |||
|- | |||
| 17. 5 ||153236||53.892||8258213||562452 | |||
|- | |||
| 18. 5||150292||144.603||21732671||462565 | |||
|} | |||
'''Fitting a Straight Line'''<br> | '''Fitting a Straight Line'''<br> | ||
[[Image:Screen_Shot_2014-04-15_at_9.51.52_PM.png]] | |||
<br> | <br> | ||
'''PCR Results Summary''' <br> | |||
Your positive control PCR result was 0.13688 μg/mL<br> | |||
Your negative control PCR result was -0.00149 μg/mL | |||
<br><br> | |||
Patient 34902 :Results were -0.0000335 μg/mL, -0.00174 μg/mL and 0.000359 μg/mL. The sample showed minimal traces of green dye. | |||
<br>Patient 39905 : Results were 0.00525 μg/mL, 0.00425 μg/mL and 0.0194 μg/mL. The sample showed traces of green dye, however, not enough to come up with a conclusion. | |||
<br><br> | |||
The final conclusion was that both patients tested negative. This was determined by comparing the concentrations to the controls. | |||
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Latest revision as of 22:58, 15 April 2014
BME 100 Spring 2014 | Home People Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3 Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6 Course Logistics For Instructors Photos Wiki Editing Help | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
OUR TEAM
LAB 5 WRITE-UPBackground InformationSYBR Green 1 Dye Single-Drop Fluorimeter How the Fluorescence Technique Works
ProcedureSmart Phone Camera Settings
Calibration For the calibration, the phone was placed into the cradle and the self-timer was readied. The flourimeter was 6 cm away from where the drop was going to be placed and the slide was placed into the holder. A sample of DNA was placed onto the slide using the micropipette. The black box was placed over the cradle and flourimeter and the picture was taken. This process was repeated for every DNA sample.
Solutions Used for Calibration
Placing Samples onto the Fluorimeter
Data AnalysisRepresentative Images of Samples
The final conclusion was that both patients tested negative. This was determined by comparing the concentrations to the controls.
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