Professional Documents
Culture Documents
Steven Buchsbaum
Point of Care (POC) refers to the ability to make a diagnosis at or near the patients location POC diagnostics has many advantages
Quicker treatment for patients Military use Food testing Environmental monitoring
It is challenging to create a POC diagnostic device that is both portable, sensitive and reliable
The IMED will be able to detect a specific target from a very low initial concentration This will be done by combining three processes on a microfluidic chip
Polymerase
The IMED is designed so that it is easy to change the target to almost anything
Microfluidics
flow
Very
This can make mixing difficult Dont need to waste expensive reagents By using pumps it is easy to automate fluid handling
Small
Easy
Polymerase Chain Reaction (PCR) is the process of exponentially duplicating DNA First described around 1971 by Kjell Kleppe but did not receive much attention In 1976 the discovery of Taq polymerase, which can withstand high temperatures, made it possible to easily automate PCR First put into practice in 1983 by Kary Mullis He received the Nobel Prize in Chemistry in 1993
Voltammetry
1. Working electrode this is where the reaction of interest is occurring, electrons flow into this electrode as reaction occurs 2. Auxiliary electrode (counter) electrons flow out of this electrode to replace leaving electrons 3. Reference electrode used to keep track of what the applied voltage
ssDNA is important because it is easy to detect It is produced using an enzyme that will digest a strand of DNA if a phosphorus is attached to the end Forward primer 5 is phosphorolated
P Replase
Blue circle is methalyne blue, a redox indicator which transfers an electron to the gold surface when a potential is applied
The IMED
Data
There is a very clear signal drop for the sample with an initial concentration of 100aM of genomic salmonella DNA while the negative control shows no signal drop. Limit of detection is around 10 aM or about 300 copies of DNA template (data now shown).
Conclusion
The
It
Continued Work
Starting
sample
Integrated Signal
on chip mixing
on detection
idea is to lyse (break apart) all cells in the blood to expose all DNA DNA is then captured using negatively charged magnetic beads These beads are then captured with a magnet DNA is removed from beads
DNA Purification
On Chip Mixing
Top of channel
Bottom of channel
Saw-tooth
Top view
Top view
Signal on Detection
Forward
Primer 5 is phosphorolated
Post PCR product
Replase
Signal on Detection
Before Hybridization: After Hybridization:
Au Electrode
Small signal:
Signal:
NO fluid flow
Acknowledgments
Scott Ferguson Kuangwen Hsieh Jonathan Adams Professor Tom Soh
Sources
Chemical Principals, Steven Zumdahl, 2005 Some Redox Indicators, L. Michaels and H. Eagle, 1930 Wikipedia, PCR diagram Invitrogen website, chargeswitch bead technology Wikipedia, Electrochemistry diagram
Questions
1) What is the polymerase chain reaction (PCR)?
a) A mixing method used when dealing with laminar flow b) A process that exponentially duplicates a DNA sequence c) The reaction that takes place during voltammetry d) A chemical reaction used to heat the IMED chip
2) The small scale of the IMED results in laminar fluid flow which makes which of the following challenging?
a) On chip fluid handling b) Predicting fluid flow patterns c) Observing the fluids d) On chip fluid mixing
Fabrication
On Chip PCR
Terms
Plasma