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Getting Started with
In-Vitro Blood Vessel
Research
Getting Started with In-Vitro Blood Vessel Research
1. History of the Pressurized Arteriograph (Gerry Herrera)
2. Anatomy of an Arteriograph System (Gerry Herrera)
3. Introduction to Research Applications and Methodology (Scott Earley)
4. Getting Started: (Scott Earley)
• KCl and PE induced constriction
• ACh-induced dilation
• Myogenic constriction
5. Tips & Troubleshooting (Scott Earley)
6. Q&A Session (Scott Earley & Gerry Herrera)
InsideScientific is an online educational environment
designed for life science researchers. Our goal is to aid in
the sharing and distribution of scientific information
regarding innovative technologies, protocols, research
tools and laboratory services.
Getting Started with In-Vitro
Blood Vessel Research
Gerry Herrera, PhD
President,
Catamount Research &
Development Inc.,
Living Systems Instrumentation
Copyright 2015 InsideScientific & Living Systems Instrumentation. All Rights Reserved.
The Halpern/Mulvany Wire Myograph
Pioneering The Way…
The Halpern/Mulvany Wire Myograph
Pioneering The Way…
The Halpern/Mulvany Wire Myograph
Pioneering The Way…
The First Arteriograph for In Vitro Vessel Research
The First Arteriograph for In Vitro Vessel Research
Automating Data Collection: Edge Detection
Automating Data Collection: Edge Detection
Automating Data Collection: Edge Detection
Anatomy of a Pressurized Arteriograph System
To be modified…
Constant Pressure
• The vast majority of
applications are well-served by
constant pressure, no
intraluminal flow setup
• Distal end of vessel is occluded
• Key components include a
pressure source, vessel
chamber, means for
temperature control, and data
acquisition hardware/software
Perfused Resistance Vessels
Advanced Application:
• It is also possible to measure vascular
responses at either constant or
variable intraluminal flow rate
Anatomy of a Pressurized Arteriograph System
To be modified…
Constant Pressure
& Flow
• Setup for intravascular flow is a bit
more complex
• Distal end of vessel remains open
• In addition to the basic items for
regulating intravascular pressure,
you will also need a pump to control
intraluminal solution flow and a way
to monitor pressure on both sides of
the cannulated vessel
Shown with optional thermistor (THRS)
• Vessel is mounted
between the cannulae
and tied in place with
nylon thread (provided
with the chamber)
• Blood is flushed out of
the vessel
• Superfusion of desired
buffer into chamber and
into vessel
Parts of a Pressure Arteriograph
Click Here for
Chamber Information
Additional Information on Equipment Required for
Setting up a Pressurized Arteriograph System
• Vessel Chambers
• Specialty Chambers
• Myographs
• Pressure Control
• Flow Control
• Video Analyzers
• Temperature Control
• Aeration & Oxygenation
• pH Measurements
• Dissection Tools
• Microscopes
• Data Acquisition
Getting Started with In-Vitro
Blood Vessel Research
Scott Earley, PhD
Associate Professor of
Pharmacology,
University of Nevada School of
Medicine
Copyright 2015 InsideScientific & Scott Earley. All Rights Reserved.
What kinds of studies can we do using Pressurized
Arteriograph Systems?
1. Smooth Muscle Cell
Contractility
2. Endothelial Function
3. Vessel Structure:
Wall Thickness & Remodeling
1. Basic Equipment and Procedures
2. Example Experiments:
• KCl- and PE-induced constriction of mouse mesenteric arteries
• ACh-induced dilation of mouse mesenteric arteries
• Myogenic constriction - mouse cerebral parenchymal
arterioles
3. Tips and Troubleshooting
What are we going to cover today?
How To Get Started
• MOPS buffered Saline
• Physiological Saline
Solution (PSS)
• High K+ PSS
• Ca2+-free PSS
• Prepare all solutions the
day of the experiment
Prepare the Solutions
Physiological Solutions
Recipes for solutions that Dr. Earley uses can be found in his
publications. Here, we provide a few saline formulations that can be
useful specifically for cases in which you cannot bubble your saline
solution – for example, when you need to keep the bath solution stagnant
without superfusion to minimize amounts of expensive drugs being added.
In most other cases, you should use a bicarbonate buffered saline in which
you bubble the saline with at CO2 mixture to get pH control. There are
many recipes for bicarbonate buffered saline in the literature.
Physiological Solutions
The following saline formulations are buffered by HEPES, including the HEPES-bicarbonate recipe.
These solutions do not require bubbling.
• HEPES-PSS Solution: This is a general purpose saline.
• High K+ (40 mM) HEPES: This is a saline in which K+ concentration has been increased to ~40 mM by
equimolar substitution with Na+. Use this for eliciting K+-induced vasoconstrictions. Some labs increase
K+ to 60 mM. If you do this, make sure to use equimolar substitution of K+ and Na+ to avoid osmolalility
problems.
• 0 Ca2+ HEPES: This is a saline used to monitor the passive responses of the vessel. Ca2+ has been
removed and a Ca2+ chelator (EGTA) has been added.
• HEPES-Bicarbonate Solution: This is a general purpose saline. Many labs prefer to have bicarbonate
present in the saline to maintain cellular bicarbonate transporter function. If you use this solution as a
standard solution, you can also prepare High K+ and 0 Ca2+ versions.
HEPES-PSS Solution
Compound M.W. mM g/L
NaCl 58.44 141.9 8.29
KCl 74.55 4.7 0.35
MgSO4 (7H2O) 246.28 1.7 0.42
EDTA 292.25 0.5 0.15
CaCl2 (2H2O) 147.02 2.8 0.41
HEPES 238.30 10.0 2.38
KH2PO4 136.09 1.2 0.16
Glucose 180.16 5.0 0.90
Titrate to pH 7.4 with 10 N NaOH. Prepare fresh daily.
(This buffer does not require bubbling)
High K+ (40 mM) HEPES
Compound M.W. mM g/L
NaCl 58.44 95.8 5.60
KCl 74.55 38.0 2.84
MgSO4 (7H2O) 246.28 1.7 0.42
EDTA 292.25 0.5 0.15
CaCl2 (2H2O) 147.02 2.8 0.41
HEPES 238.30 10.0 2.38
KH2PO4 136.09 1.2 0.16
Glucose 180.16 5.0 0.90
Use this equimolar substitution of KCl and NaCl. All other reagent amounts stay the same. Titrate to pH 7.4 with 10 N NaOH .
Prepare fresh daily.
(This buffer does not require bubbling)
0 Ca2+ HEPES
Compound M.W. mM g/L
NaCl 58.44 141.9 8.29
KCl 74.55 4.7 0.35
MgSO4 (7H2O) 246.28 1.7 0.42
EDTA 292.25 0.5 0.15
EGTA 380.40 2.0 0.75
HEPES 238.30 10 2.38
KH2PO4 136.09 1.2 0.16
Glucose 180.16 5.0 0.90
This formula is Identical to HEPES-PSS, except CaCl2 is replaced by EGTA. Titrate to pH 7.4 with 10 N NaOH . Prepare fresh daily.
(This buffer does not require bubbling)
HEPES-Bicarbonate Solution (This buffer does not require bubbling)
Compound M.W. mM g/L
NaCl 58.44 130.00 7.60
KCl 74.55 4.00 0.30
MgSO4 (7H2O) 246.28 1.20 0.30
NaHCO3 84.00 4.00 0.34
CaCl2 (2H2O) 147.02 1.80 0.27
HEPES 238.30 10.0 2.38
KH2PO4 136.09 1.18 0.16
Glucose 180.16 6.00 1.08
EDTA 292.25 0.03 0.009
Osmolality before adding glucose is approximately 300 mOsm/L. Titrate to pH 7.4 with 10 N NaOH . Prepare fresh daily.
Isolating the Vessels
• Microscope
• Dissection dishes
• Dissection tools
Mesentery Brain
Isolating the Vessels
• Mesentery pinned out
and fat removed
• Veins and Arteries
Isolating the Vessels
• We weren’t able to come
up with any images for
this – Gerry, do you have
anything?
• ….
• ….
• ….
Making Vessel Ties
Mounting Vessels in the Pressure Myograph
Making Vessel Ties:
• Use multi-filament braided nylon fibers
• Cut to desired length (~5-10mm)
• Tease individual filaments apart –
use 1 filament per tie
• Make several ties and store them in a
covered petri-dish using double-sided
tape
• Pick up tie from end attached to tape,
NOT from the free end
• We weren’t able to come
up with any images for
this – Gerry, do you have
anything?
• ….
• ….
• ….
Attach proximal end
& flush out blood…
Attach distal end.
Mounting Vessels in the Pressure Myograph
Transfer Chamber to
Recording System
• Place chamber on the
microscope stage
• Connect inlet and outlet
superfusion
• Connect pressure
control system
Keeping the Vessels Alive and Happy
Intraluminal
Pressure
Temperature Flow Rate pH
Follow links for additional information on suggested equipment
Collecting Data
• Edge Detection
• Data Acquisition
System
The Complete System
Example Experiments
Example 1: KCl-Induced Constriction
• Expose the vessel to elevated
extracellular KCl (60 mM) – High
K+ PSS.
• Collapses the K+ gradient,
resulting in depolarization of the
smooth muscle cell plasma
membrane.
• Ca2+ influx through L-type Ca2+
channels causes vascular smooth
muscle contraction and
vasoconstriction.
K+
K+ K+
K+ K+K+
Ca2+
SMC
Vessel superfused with High K+ PSS:
Baseline Diameter High K+ PSS
Example 2: PE-Concentration Response Curve
• Phenylephrine (PE) binds to
the type-1 adrenergic
receptor (α1 receptor) on the
smooth muscle cell plasma
membrane.
• This causes an increase in
intracellular Ca2+, resulting in
smooth muscle cell
contraction and
vasoconstriction.
PE
Ca2+
SMC
10-9 M PE 10-8 M PE 10-7 M PE 10-6 M PE 10--5 M PE
Vessel superfused with increasing concentrations of PE:
Effects of increasing PE concentration on vessel diameter:
• Recording of
luminal diameter
versus time as
increasing
concentrations of
PE are delivered
• When doing dose
response be sure to
allow the recording
to stabilize
Plot the Data…
10-9 10-8 10-7 10-6 10-5 10-4 10-3
0
20
40
60
[PE]
%Constriction
n=5
% Constriction
=
(Baseline Diameter – Diameter with PE)
Baseline Diameter
Example 3: ACh-Induced Relaxation (Endothelium Viability)
• Acetylcholine (ACh) binds to
muscarinic type-2 (M2) receptors
on the endothelial cell plasma
membrane.
• This causes the release of NO and
EDHF.
• These factors diffuse to the
underlying vascular smooth muscle
cells to decrease intracellular Ca2+
and cause dilation.
• Gold standard for intact endothelial
function.
Endothelial
cell
SMC
ACh
Ca2+
NO
EDHF
Baseline Diameter Pre-Constrict: 10-6 M PE 10-4 M ACh + 10-6 M PE
Example 3: ACh-Induced Relaxation (Endothelium Viability)
ACh
Example 3: ACh-Induced Relaxation (Endothelium Viability)
• Vessel was pre-constricted
with PE
• ACh was applied at the peak
of the steady-state PE-
constriction
Damaging the endothelium
Perfuse Vessel With AirBaseline Diameter
Following Endothelial Cell Damage:
Baseline diameter Pre-constrict 10-6 M PE 10-4 M ACh + 10-6 M PE
Plot the Data…
10-10 10-9 10-8 10-7 10-6 10-5 10-4 10-3 10-2
0
50
100
150
[ACh]
%Vasodilation
Endothelium Intact
Endothelium Damaged
n=5
%Vasodilation
=
% Reversal of Preconstriction
=
[(Baseline – PE+ACh)/Baseline] –
[(Baseline – PE/Baseline)]
[(Baseline – PE)/Baseline]
Example 4: Myogenic Response – Cerebral Parenchymal Arterioles
• Record diameter during step-
wise increases in intraluminal
pressure
• Repeat while superfusing
vessels with Ca2+-free PSS to
determine the passive
diameter
• %Myogenic tone = [(Passive
Diameter – Active
Diameter)/Passive Diameter]
P (mmHg)
passive
active
• The learning curve – do the procedure in examples
1-3 (KCl-induced constriction, PE-concentration response curve,
ACh-induced dilation) using second order mesenteric arteries until
you get reproducible results.
• Work fast- remove organs and isolate vessels as quickly as possible
• Make solutions fresh each day.
• Stretching the vessel in the initial prep
• pH/O2/temperature
Tips & Pointers
Thank You!
For additional information on Pressure Arteriograph
Systems, Wire Myographs, and solutions for
studying vessel function in-vitro visit:
http://www.livingsys.com/index.html

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Getting Started with In-Vitro Blood Vessel Research

  • 1. Getting Started with In-Vitro Blood Vessel Research
  • 2. Getting Started with In-Vitro Blood Vessel Research 1. History of the Pressurized Arteriograph (Gerry Herrera) 2. Anatomy of an Arteriograph System (Gerry Herrera) 3. Introduction to Research Applications and Methodology (Scott Earley) 4. Getting Started: (Scott Earley) • KCl and PE induced constriction • ACh-induced dilation • Myogenic constriction 5. Tips & Troubleshooting (Scott Earley) 6. Q&A Session (Scott Earley & Gerry Herrera)
  • 3. InsideScientific is an online educational environment designed for life science researchers. Our goal is to aid in the sharing and distribution of scientific information regarding innovative technologies, protocols, research tools and laboratory services.
  • 4. Getting Started with In-Vitro Blood Vessel Research Gerry Herrera, PhD President, Catamount Research & Development Inc., Living Systems Instrumentation Copyright 2015 InsideScientific & Living Systems Instrumentation. All Rights Reserved.
  • 5. The Halpern/Mulvany Wire Myograph Pioneering The Way…
  • 6. The Halpern/Mulvany Wire Myograph Pioneering The Way…
  • 7. The Halpern/Mulvany Wire Myograph Pioneering The Way…
  • 8. The First Arteriograph for In Vitro Vessel Research
  • 9. The First Arteriograph for In Vitro Vessel Research
  • 10. Automating Data Collection: Edge Detection
  • 11. Automating Data Collection: Edge Detection
  • 12. Automating Data Collection: Edge Detection
  • 13. Anatomy of a Pressurized Arteriograph System To be modified… Constant Pressure • The vast majority of applications are well-served by constant pressure, no intraluminal flow setup • Distal end of vessel is occluded • Key components include a pressure source, vessel chamber, means for temperature control, and data acquisition hardware/software
  • 14. Perfused Resistance Vessels Advanced Application: • It is also possible to measure vascular responses at either constant or variable intraluminal flow rate
  • 15. Anatomy of a Pressurized Arteriograph System To be modified… Constant Pressure & Flow • Setup for intravascular flow is a bit more complex • Distal end of vessel remains open • In addition to the basic items for regulating intravascular pressure, you will also need a pump to control intraluminal solution flow and a way to monitor pressure on both sides of the cannulated vessel
  • 16. Shown with optional thermistor (THRS) • Vessel is mounted between the cannulae and tied in place with nylon thread (provided with the chamber) • Blood is flushed out of the vessel • Superfusion of desired buffer into chamber and into vessel Parts of a Pressure Arteriograph Click Here for Chamber Information
  • 17. Additional Information on Equipment Required for Setting up a Pressurized Arteriograph System • Vessel Chambers • Specialty Chambers • Myographs • Pressure Control • Flow Control • Video Analyzers • Temperature Control • Aeration & Oxygenation • pH Measurements • Dissection Tools • Microscopes • Data Acquisition
  • 18. Getting Started with In-Vitro Blood Vessel Research Scott Earley, PhD Associate Professor of Pharmacology, University of Nevada School of Medicine Copyright 2015 InsideScientific & Scott Earley. All Rights Reserved.
  • 19. What kinds of studies can we do using Pressurized Arteriograph Systems? 1. Smooth Muscle Cell Contractility 2. Endothelial Function 3. Vessel Structure: Wall Thickness & Remodeling
  • 20. 1. Basic Equipment and Procedures 2. Example Experiments: • KCl- and PE-induced constriction of mouse mesenteric arteries • ACh-induced dilation of mouse mesenteric arteries • Myogenic constriction - mouse cerebral parenchymal arterioles 3. Tips and Troubleshooting What are we going to cover today?
  • 21. How To Get Started
  • 22. • MOPS buffered Saline • Physiological Saline Solution (PSS) • High K+ PSS • Ca2+-free PSS • Prepare all solutions the day of the experiment Prepare the Solutions
  • 23. Physiological Solutions Recipes for solutions that Dr. Earley uses can be found in his publications. Here, we provide a few saline formulations that can be useful specifically for cases in which you cannot bubble your saline solution – for example, when you need to keep the bath solution stagnant without superfusion to minimize amounts of expensive drugs being added. In most other cases, you should use a bicarbonate buffered saline in which you bubble the saline with at CO2 mixture to get pH control. There are many recipes for bicarbonate buffered saline in the literature.
  • 24. Physiological Solutions The following saline formulations are buffered by HEPES, including the HEPES-bicarbonate recipe. These solutions do not require bubbling. • HEPES-PSS Solution: This is a general purpose saline. • High K+ (40 mM) HEPES: This is a saline in which K+ concentration has been increased to ~40 mM by equimolar substitution with Na+. Use this for eliciting K+-induced vasoconstrictions. Some labs increase K+ to 60 mM. If you do this, make sure to use equimolar substitution of K+ and Na+ to avoid osmolalility problems. • 0 Ca2+ HEPES: This is a saline used to monitor the passive responses of the vessel. Ca2+ has been removed and a Ca2+ chelator (EGTA) has been added. • HEPES-Bicarbonate Solution: This is a general purpose saline. Many labs prefer to have bicarbonate present in the saline to maintain cellular bicarbonate transporter function. If you use this solution as a standard solution, you can also prepare High K+ and 0 Ca2+ versions.
  • 25. HEPES-PSS Solution Compound M.W. mM g/L NaCl 58.44 141.9 8.29 KCl 74.55 4.7 0.35 MgSO4 (7H2O) 246.28 1.7 0.42 EDTA 292.25 0.5 0.15 CaCl2 (2H2O) 147.02 2.8 0.41 HEPES 238.30 10.0 2.38 KH2PO4 136.09 1.2 0.16 Glucose 180.16 5.0 0.90 Titrate to pH 7.4 with 10 N NaOH. Prepare fresh daily. (This buffer does not require bubbling)
  • 26. High K+ (40 mM) HEPES Compound M.W. mM g/L NaCl 58.44 95.8 5.60 KCl 74.55 38.0 2.84 MgSO4 (7H2O) 246.28 1.7 0.42 EDTA 292.25 0.5 0.15 CaCl2 (2H2O) 147.02 2.8 0.41 HEPES 238.30 10.0 2.38 KH2PO4 136.09 1.2 0.16 Glucose 180.16 5.0 0.90 Use this equimolar substitution of KCl and NaCl. All other reagent amounts stay the same. Titrate to pH 7.4 with 10 N NaOH . Prepare fresh daily. (This buffer does not require bubbling)
  • 27. 0 Ca2+ HEPES Compound M.W. mM g/L NaCl 58.44 141.9 8.29 KCl 74.55 4.7 0.35 MgSO4 (7H2O) 246.28 1.7 0.42 EDTA 292.25 0.5 0.15 EGTA 380.40 2.0 0.75 HEPES 238.30 10 2.38 KH2PO4 136.09 1.2 0.16 Glucose 180.16 5.0 0.90 This formula is Identical to HEPES-PSS, except CaCl2 is replaced by EGTA. Titrate to pH 7.4 with 10 N NaOH . Prepare fresh daily. (This buffer does not require bubbling)
  • 28. HEPES-Bicarbonate Solution (This buffer does not require bubbling) Compound M.W. mM g/L NaCl 58.44 130.00 7.60 KCl 74.55 4.00 0.30 MgSO4 (7H2O) 246.28 1.20 0.30 NaHCO3 84.00 4.00 0.34 CaCl2 (2H2O) 147.02 1.80 0.27 HEPES 238.30 10.0 2.38 KH2PO4 136.09 1.18 0.16 Glucose 180.16 6.00 1.08 EDTA 292.25 0.03 0.009 Osmolality before adding glucose is approximately 300 mOsm/L. Titrate to pH 7.4 with 10 N NaOH . Prepare fresh daily.
  • 29. Isolating the Vessels • Microscope • Dissection dishes • Dissection tools
  • 31. • Mesentery pinned out and fat removed • Veins and Arteries Isolating the Vessels
  • 32. • We weren’t able to come up with any images for this – Gerry, do you have anything? • …. • …. • …. Making Vessel Ties Mounting Vessels in the Pressure Myograph Making Vessel Ties: • Use multi-filament braided nylon fibers • Cut to desired length (~5-10mm) • Tease individual filaments apart – use 1 filament per tie • Make several ties and store them in a covered petri-dish using double-sided tape • Pick up tie from end attached to tape, NOT from the free end
  • 33. • We weren’t able to come up with any images for this – Gerry, do you have anything? • …. • …. • …. Attach proximal end & flush out blood… Attach distal end. Mounting Vessels in the Pressure Myograph
  • 34. Transfer Chamber to Recording System • Place chamber on the microscope stage • Connect inlet and outlet superfusion • Connect pressure control system
  • 35. Keeping the Vessels Alive and Happy Intraluminal Pressure Temperature Flow Rate pH Follow links for additional information on suggested equipment
  • 36. Collecting Data • Edge Detection • Data Acquisition System
  • 39. Example 1: KCl-Induced Constriction • Expose the vessel to elevated extracellular KCl (60 mM) – High K+ PSS. • Collapses the K+ gradient, resulting in depolarization of the smooth muscle cell plasma membrane. • Ca2+ influx through L-type Ca2+ channels causes vascular smooth muscle contraction and vasoconstriction. K+ K+ K+ K+ K+K+ Ca2+ SMC
  • 40. Vessel superfused with High K+ PSS: Baseline Diameter High K+ PSS
  • 41. Example 2: PE-Concentration Response Curve • Phenylephrine (PE) binds to the type-1 adrenergic receptor (α1 receptor) on the smooth muscle cell plasma membrane. • This causes an increase in intracellular Ca2+, resulting in smooth muscle cell contraction and vasoconstriction. PE Ca2+ SMC
  • 42. 10-9 M PE 10-8 M PE 10-7 M PE 10-6 M PE 10--5 M PE Vessel superfused with increasing concentrations of PE:
  • 43. Effects of increasing PE concentration on vessel diameter: • Recording of luminal diameter versus time as increasing concentrations of PE are delivered • When doing dose response be sure to allow the recording to stabilize
  • 44. Plot the Data… 10-9 10-8 10-7 10-6 10-5 10-4 10-3 0 20 40 60 [PE] %Constriction n=5 % Constriction = (Baseline Diameter – Diameter with PE) Baseline Diameter
  • 45. Example 3: ACh-Induced Relaxation (Endothelium Viability) • Acetylcholine (ACh) binds to muscarinic type-2 (M2) receptors on the endothelial cell plasma membrane. • This causes the release of NO and EDHF. • These factors diffuse to the underlying vascular smooth muscle cells to decrease intracellular Ca2+ and cause dilation. • Gold standard for intact endothelial function. Endothelial cell SMC ACh Ca2+ NO EDHF
  • 46. Baseline Diameter Pre-Constrict: 10-6 M PE 10-4 M ACh + 10-6 M PE Example 3: ACh-Induced Relaxation (Endothelium Viability)
  • 47. ACh Example 3: ACh-Induced Relaxation (Endothelium Viability) • Vessel was pre-constricted with PE • ACh was applied at the peak of the steady-state PE- constriction
  • 48. Damaging the endothelium Perfuse Vessel With AirBaseline Diameter
  • 49. Following Endothelial Cell Damage: Baseline diameter Pre-constrict 10-6 M PE 10-4 M ACh + 10-6 M PE
  • 50. Plot the Data… 10-10 10-9 10-8 10-7 10-6 10-5 10-4 10-3 10-2 0 50 100 150 [ACh] %Vasodilation Endothelium Intact Endothelium Damaged n=5 %Vasodilation = % Reversal of Preconstriction = [(Baseline – PE+ACh)/Baseline] – [(Baseline – PE/Baseline)] [(Baseline – PE)/Baseline]
  • 51. Example 4: Myogenic Response – Cerebral Parenchymal Arterioles • Record diameter during step- wise increases in intraluminal pressure • Repeat while superfusing vessels with Ca2+-free PSS to determine the passive diameter • %Myogenic tone = [(Passive Diameter – Active Diameter)/Passive Diameter] P (mmHg) passive active
  • 52. • The learning curve – do the procedure in examples 1-3 (KCl-induced constriction, PE-concentration response curve, ACh-induced dilation) using second order mesenteric arteries until you get reproducible results. • Work fast- remove organs and isolate vessels as quickly as possible • Make solutions fresh each day. • Stretching the vessel in the initial prep • pH/O2/temperature Tips & Pointers
  • 53. Thank You! For additional information on Pressure Arteriograph Systems, Wire Myographs, and solutions for studying vessel function in-vitro visit: http://www.livingsys.com/index.html