FLUORESCENT DETECTION USING OPTICAL FIBERS WITH CARDIAC MYOCYTES DESIGN GROUP EIGHT FLUORESCENT DETECTION USING OPTICAL FIBERS WITH CARDIAC MYOCYTES Paul Clark, Martin Garcia, Chris Gorga, John Ling, Jordan LoRegio

Introduction  Importance of calcium excretion by cardiac myocytes  Changes in the interstitial calcium concentration can be equated to the force of myocyte concentration  Changes in calcium concentration can be visualized via optical detection of fluorescence  Quantifying the forces on a cellular level is best  Therefore we use BioMEMs

Motivation & Purpose Quantifying cellular metabolism is one of the best ways to understand the physical workings of the cell Optical Detection allows for real time data acquisition

Motivation & Purpose (con’t) Due to the prevalence of cardiac related diseases in the general population understanding cardiac function is leading scientific research Understanding cardiac cells will help when looking at other muscle cells

Goals  1 st Goal:  Create a microfluidic device to equate changes in calcium concentration and flourescence  The device must...  Incorporate an optical fiber  Infuse fluoroscein and calcium solutions (of varying concentrations)  Create a program that will assign a magnitude to the measured fluorescence value  This will be done by implementing a Lab View program  2 nd Goal:  Create a microfluidic device that incorporates a single cardiac myocyte  The device must…  Hold a single cardiac myocyte  Incorporate electrodes to induce cell contraction  Incorporate the previously created LabView program

Results – Goal #1  Master Fabrication  SU was spun onto silicon substrate until thickness of 50 microns was obtained  Master then cured in oven for 45 minutes  Device Fabrication  PDMS is mixed in a 10:1 ratio of solvent to solute  Mixed for 2 minutes in centrifuge  Micro-manipulator was used to align fiber optic cable directly over one channel of the master  PDMS then poured over master Caution was taken as to ensure there where no bubbles

Testing! ! !  The optical fiber is fitted with a connector that connects directly to the box (pictured left)  This box, designed by Tobias Meyer, is able to amplify signals and record them graphically

Status  Using the clean room to cast PDMS devices with different size optical fibers  Currently we are using a plastic 500 micron fiber because of its flexibility and resilience.  Testing the limits of detection of the different fibers  Perfecting the Lab View module…  Store recorded data to an excel file for analysis  Find proper amplification set-up

Conclusions  Experimental setup to achieve goal one is ready for testing  Data from this will show relationships between calcium concentration and magnitude of fluorescence  What’s next… Find minimum concentration of fluorescein needed Create a trend that relates calcium concentration to magnitude of fluorescence Begin design on the second device

Questions ? ? ?