explore different tubing sizes and materials integrate flow chamber with fluorescent microscope
DESCRIPTION
Design of a Flow Chamber for Measuring Cell Membrane Permeability. Laura Dennis and Matthew Coblyn OSU School of CBEE, CHE 415-416. BACKGROUND. PROCESS DESCRIPTION. PROGRESS TO DATE. - PowerPoint PPT PresentationTRANSCRIPT
• Cryopreserving intact tissue is an important biological function in the biomedical industry and is dependent on cell membrane permeability to water• There is a lack of techniques to measure cell membrane permeability, but fluorescence quenching is promising • A fluorescent dye, calcein is quenched by molecules in the cell cytoplasm• Fluorescence color intensity is observed during cell exposure to isotonic and anisotonic (salt and no salt) solutions using a fluorescence microscope
• Linear relationship between fluorescence intensity and cell volume is observed
• Design flow chamber to measure cell membrane permeability of intact tissue under strict environmental conditions• Use fluorescence quenching technique to observe cell membrane reaction to anisotonic and isotonic solutions• Fluorescence intensity increases upon cell swelling and decreases upon cell shrinkage We thank Dr. Adam Higgins for sponsoring the project, Dr. Phillip Harding for
guidance, HaiYue Han for helping with SolidWorks, Steve Adams, William Murray and his students, and Andrew Brickman for fabricating the flow chamber, and Dr. AlexandreYokochi for his helpful suggestions.
1. Explore different tubing sizes and materials2. Integrate flow chamber with fluorescent
microscope3. Vary temperature of solutions passing over the
cells to mimic cooling conditions
BACKGROUND
Isotonic Hypertonic
DESIGN OF A FLOW CHAMBER FOR MEASURING CELL MEMBRANE PERMEABILITY Laura Dennis and Matthew Coblyn
OSU School of CBEE, CHE 415-416
PROCESS DESCRIPTION
PURPOSE AND HYPOTHESISFUTURE WORK
ACKNOWLEDGEMENTS
Gasket
Glass coverslip with cultured cells
UV lightsource and microscope
Cross-section of flow chamber
A salt solution is pumped from syringe to flow chamber housing. It then enters the heat exchanger, which consists of 27 cm of 1/16” Tygon tubing bathed in heating/cooling fluid which is continuously circulated with the water bath source. The salt solution immediately enters the flow chamber after exiting the heat exchanger.
Water bath
Flow chamber
Thermocouple reader
Syringe Pump
Side view of chamber
Create salt concentration profiles that model a step change in concentration by eliminating the inlet tubing preceding the flow chamber that both salt solutions must flow through, thereby minimizing mixing.
PROGRESS TO DATE
Figure 1 Cell swelling and cell shrinking as a function of exposure to isotonic and anisotonic solutions, respectively.
Figure 3 Effects of hi, ho, and ktubing on the overall heat transfer coefficient.
Figure 2 Experimental values are plotted along with theoretically expected trends. Flow rate refers to the tube side flow. High and low pump settings refer to the water bath pump that flows fluid through the shell side. Tube 1 and tube 2 refer to the two separate tubes within the heat exchanger that feed the flow chamber different solutions
Nu1.86 RePrDL
13hDk fluid
0 50 100 150 200 250 300 350 4000
20
40
60
80
100
120
140
High pump setting, tube 1
High pump setting, tube 2
Low pump setting, tube 1
Theoretical, Low pump setting
Theoretical, High pump setting
Flowrate (ml/hr)
U (W
/m2-
K)
0.1 1 100
50
100
150
200
250
300
350
400
450
ho hi k
Factor change from theoretical value
U (W
/m2-
K)
ln( / )1 12
i o i i
i tubing o o
D D D DU h k D h
Re D
Pr CPk fluid