Design Group Eight Fluorescent Detection using Optical Fibers with Cardiac Myocytes

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Design Group Eight Fluorescent Detection using
Optical Fibers with Cardiac Myocytes

• Paul Clark, Martin Garcia, Chris Gorga, John
  Ling, Jordan LoRegio

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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

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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

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Motivation Purpose (cont)

• 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

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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

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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 101 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

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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

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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

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Conclusions

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

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Questions ? ? ?