Download Oral Report IV- March 15, 2007

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