DESIGN GROUP EIGHT FLUORESCENT DETECTION USING FLUORESCENT DETECTION USING OPTICAL FIBERS WITH CARDIAC OPTICAL FIBERS WITH CARDIAC MYOCYTESMYOCYTES

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 1st 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

2nd 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-8 2025 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 ? ? ?