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Phosphorescence O2 Measurement System
PMOD 2000
The PMOD-2000 uses a noninvasive optical method for detecting the
concentration of O2 in tissue. The operating principle for the unit is
based on the quenching of phosphorescence by oxygen in phosphors
that have been injected into the blood. To determine the concentration
of O2 in the area of tissue being measured, phosphorescence
lifetime measurements are taken using a sinusoidally modulated light
source for excitation and frequency lock for phosphorescence detection.
The PMOD operates over a frequency range from 200 to 20,000
Hz, allowing measurements of phosphorescence lifetimes from <15
microseconds to 1.5 milliseconds. Measurements can be carried out
either at constant frequency or constant phase shift. This technique
differs from other methods of O2 detection in that this method is totally
noninvasive, with the fiber optic bundle outside of the tissue in
which the O2 is being measured. The PMOD-2000 has a very high
accuracy throughout the physiological range and provides real time
measurements with high temporal resolution (~1 sec/measurement).
The phosphors that are used with this system have absolute
calibration and follow Stern-Volmer behavior. Once a phosphor is
calibrated for a given temperature and solvent conditions these values
are stable, eliminating the possibility of measurement drift and
the requirement for re-calibration at the time of the measurement.
The system has two channels (sources) of excitation light. The standard
system comes with one wavelength set to 524 nm (suitable for the
porphyrin based phosphors) and 635 nm (suitable for the tetrabenzoporphyrin
based phosphors). Other combinations of excitation wavelengths
are available. Please call our technical sales department for further
information.
Phosphorescence emission is in the range of 660 to 760 nm (porphyrins)
or 760 to 900 nm (tetrabenzoporphyrins). Since bright ambient
light can saturate the detector, it is recommended that the work
area be lighted with DC powered monochromatic light sources.
The 慍ool Colors?LED lamps are specifically designed for this purpose.
When an appropriate lamp is used, this light does not interfere
with the phosphorescence measurements.
Types of Phosphors and Phosphor Kits Available
Pd-meso-tetra (4-carboxyphenyl) porphyrin
This phosphor (Oxyphor RO) is dissolved in the presence of 0.5% or
greater of bovine serum albumin. It provides excellent oxygen sensitivity
and has been accurately calibrated (see Lo et al., 1996). This
phosphor has an absorption band near 524 nm and emission near
690 nm and can be used for surface O2 measurements in tissues.
Lyophilized Porphyrin Kit*
1.2 g. approx. amount needed for approx. 30 rat (rats weighing
300 g each) and 10 cat (cats weighing 3 kg each) experiments using
the PMOD instrument. Lyophilized porphyrin contains (approx.):
9% Pd-meso-tetra (4-carboxyphenyl) porphine which is bound to
80% BSA (clinical grade, low fatty acid), 8% NaCl, and 3% TRISMA
buffer. Accessories include one (1) 10 cc syringe, three (3) syringe
filters (25 x .22 micron), and three (3) 4.0 mL amber glass vials with
foil caps.
Pd-meso-tetra (4-carboxyphenyl) porphyrin dendrimer
Oxyphor R2: This new water soluble phosphor has significant advantages
over the parent compound for experiments in vivo. The quenching
properties are similar to those of the parent compound with
respect to the effect of albumin. However, because it is highly water
soluble it can be injected into the blood as a simple solution in physiological
saline. The pH dependence of Oxyphor R2 is also very small
over the physiological range (see Lo et al., 1997). Oxyphor R2 is
largely excreted in the urine in mice and other animals. In mice,
excretion occurs with a half time of less than 2 hours.
Pd-tetra-(4-carboxyphenyl) tetrabenzoporphyrin dendrimer
Oxyphor G2: This is a water soluble phosphor that has an absorption
band near 636 nm and emission near 800 nm and can be used for O2
measurements in tissue up to 3 cm thick.
BS4 69-0502
?Two built-in light sources; one for surface (0.5 to 1 mm) and
one for deep tissue (2 to 3 cm) measurements
?Noninvasive oxygen measurements in tissue in-vivo or in-vitro
?Linear relationship between 1/lifetime and pO2
?Stable over time, no sensitivity or baseline drift
?Absolute calibration; calibrate only once
?Windows operating environment
?Temporal resolution < 1.2 sec/measurement
?Evaluate a wide range of tissue physiology and pathology
?Vascular and cardiopulmonary function
?Tumor oxygenation
?Status of skin grafts
?Diabetic vascular complications
?Ischemia/reperfusion injury, etc.
?Visible and near infrared phosphors for different tissue depth
measurements
?Measurements in-vitro in sealed glass chambers, no oxygen
exchange with the environment
11,000 Specialty Products to Enhance Your Bioresearch M8
Dissolved O2
Specialized Tools For Bioresearch
Catalog No. $ Product
BS4 69-0502 10,910.00 PMOD-2000 Phosphorescence Oxygen
Measurement System
BS4 69-0503 70.00 Cool Color Lamp, Amber (592 nm maximal intensity),
115 VAC, 60 Hz
BS4 69-0504 70.00 Cool Color Lamp, Amber (592 nm maximal intensity),
230 VAC, 50 Hz
BS4 69-0505 70.00 Cool Color Lamp, Red (638 nm maximal intensity),
115 VAC, 60 Hz
BS4 69-0506 70.00 Cool Color Lamp, Red (638 nm maximal intensity),
230 VAC, 50 Hz
BS4 69-0507 85.00 Cool Color Lamp, Green (524 nm maximal intensity),
115 VAC, 60 Hz
BS4 69-0508 85.00 Cool Color Lamp, Green (524 nm maximal intensity),
230 VAC, 50 Hz
BS4 69-0509 322.00 Phosphor, Pd-meso-tetra (4-carboxyphenyl)
porphyrin, 1 gm Vial (Oxyphor RO)
BS4 69-0510 791.00 Phosphor, Pd-meso-tetra (4-carboxyphenyl) porphyrin
dendrimer, 1 gm Vial (Oxyphor R2)
BS4 69-0511 315.00 Phosphor, Pd-tetra (4-carboxyphenyl)
tetrabenzoporphyrin dendrimer (Oxyphor G2), 20 mg
BS4 72-6180 355.00 Phosphor, Pd-tetra (4-carboxyphenyl)
tetrabenzoporphyrin dendrimer (Oxyphor G2), 50 mg
BS4 72-6181 599.00 Phosphor, Pd-tetra (4-carboxyphenyl)
tetrabenzoporphyrin dendrimer (Oxyphor G2), 100 mg
Specifications
Detection Range Air Saturation to < 0.1 Torr
Sensitivity 1 part in 103
Temporal Resolution Minimum 20 msec with measurements repeatable
each 1.2 sec
Temperature Dependence Approximately 3% per degree Celsius
Power Wall Transformer 100/230 VAC, 50/60 Hz
Dimensions 26 x 33.7 x 14 cm (10.25 x 13.25 x 5.5 in)
Weight 5 kg (11 lbs)
The 慍ool Colors?LED lamps are specifically designed for this purpose.
When an appropriate lamp is used, this light does not interfere
with the phosphorescence measurements.
The Windows compatible software included with the PMOD-2000 system
is used to configure the system抯 parameters and to display and
store the measurement frequency, signal amplitude, phase, and time
of measurement as well as the oxygen pressure. The time course of
phosphorescence lifetime or oxygen pressure can be continuously
plotted on screen as the data are taken. All of the displayed signal
parameters can be saved to named files for further analysis etc. The
files are in ASCII and are easily read by other programs such as
Excel? Sigma Plot? etc. Systems come complete with a 6 mm OD
bifurcated optical fiber cable. Other optical fiber configurations are
also available, please call our technical sales department for additional
information.
Computer Requirements: The operating software is for DOS or
Windows 95/98/NT and requires an IBM compatible PC. Only the
Windows NT version will allow determination of the distribution of
phosphorescence lifetimes. In any case, communication with the
computer requires a 16550 UART serial port (or later).
Method Reference Publications:
1) Vanderkooi, J.M., Maniara, G., Green, T.J., and Wilson, D.F. An optical method for measurement
of dioxygen concentration based on quenching of phosphorescence. J. Biol.
Chem. 262: 5476-5482, 1987.
2) Wilson, D.F., Rumsey,W.L., Green, T.J., and Vanderkooi, J.M. (1988) The oxygen dependence
of mitochondrial oxidative phosphorylation measured by a new optical method
for measuring oxygen. J. Biol. Chem. 263: 2712-2718.
3) Robiolio, M., Rumsey,W.R. and Wilson, D.F. (1989) Oxygen diffusion and mitochondrial
respiration in neuroblastoma cells. Amer. J. Physiol., 256: C1207-C1213.
4) Shonat, R.D.,Wilson, D.F., Riva, C.E., and Pawlowski, M. (1992) Oxygen distribution in
the retinal and choroidal vessels of the cat as measured by a new phosphorescence
imaging method. Applied Optics 31, 3711-3718.
5) Vinogradov, S.A. and Wilson, D.F. (1995) Metallotetrabenzoporphyrins. New phosphorescent
probes for oxygen measurements. J. Chem. Soc. Perkin Trans. II, 2, 103-111.
6) Vinogradov, S.A., Lo, L.-W., Jenkins, W.T., Evans, S.M., Koch, C., and Wilson, D.F. (1996)
Non invasive imaging of the distribution of oxygen in tissue in vivo using near
infrared phosphors. Biophys. J. 70, 1609-1617.
7) Lo, L.-W., Koch, C.J., and Wilson, D.F., (1996) Calibration of oxygen dependent quenching
of the phosphorescence of Pd-meso-tetra (4-carboxyphenyl) porphine: a phosphor
with general application for measuring oxygen concentration in biological systems.
Analy. Biochem. 236, 153-160.
8) Lo, L.-W.,Vinogradov, S.A., Koch, C.J., and Wilson, D.F. A new, water soluble, phosphor for
oxygen measurements in vivo. Adv. Exptl. Med. Biol. 428, 651-656, 1997.
9) Vinogradov, S.A., Lo, L.-W., and Wilson, D.F., (1999) Dendritic polyglutamic porphyrins:
probing porphyrin protection by oxygen dependent quenching of phosphorescence.
Chem. Europ. J., 5 (4), 1388-1347.
Selected Application Reports: (selected from more than 60)
1) Yonetani, M., Huang, Ch-Ch., Lajevardi, N., Pastuszko, A., Delivoria-Papadopoulos,
M.,and Wilson, D.F. (1994) Effect of hemorrhagic hypotension on extracellular level of
dopamine, cortical oxygen pressure and blood flow in brain of newborn piglets.
Neuroscience Letters, 180, 247-252.
2) Rumsey, W.L., Pawlowski, M., Lejavardi, N., and Wilson, D.F. (1994) Oxygen pressure
distribution in the heart in vivo and evaluation of the ischemic 揵order zone? Am. J.
Physiol. 266: H1676-1680.
3) Huang, Ch-Ch., Yonetani, M., Lajevardi, N., Delivoria-Papadopoulos, M., Pastuszko, A.
and Wilson D.F. (1995) Comparison of post-asphyxial resuscitation with 100% and
21% oxygen on striatal dopamine metabolism in newborn piglets. J. Neurochemistry,
64, 292-298.
4) Song, D., Olano, M., Wilson, D.F., Pastuszko, A., Tammela, O., Nho, K., and Shorr, R.G.L.
(1995) Comparison of the efficacy of blood and polyethylene glycol-hemoglobin in
recovery of newborn piglets from hemorrhagic hypotension: effect on blood pressure,
cortical oxygen, and extracellular dopamine in the brain. Transfusion 35, 552-558.
5) Poole, D.C.,Wagner, P.D., and Wilson, D.F. (1995) Diaphragm microvascular plasma PO2
measured in vivo. J.Appl. Physiol. 79(6), 2050-2057.
6) Olano, M., Song, D., Murphy, S.,Wilson, D.F., and Pastuszko, A. (1995) Relationships of
dopamine, cortical oxygen pressure, and hydroxyl radicals in brain of newborn piglets
during hypoxia and posthypoxic recovery. J. Neurochem. 65, 1205-1212.
7) Cerniglia, G.J., Wilson, D.F., Pawlowski, M., Vinogradov, S.A., and Biaglow, J. (1997)
Intravascular oxygen distribution in subcutaneous 9L tumors and radiation sensitivity.
J.Appl. Physiol. 82(6), 1939-1945.
8) Rumsey,W.R., Abbott, B., Lo, L.-W., Vinogradov, S.A., and Wilson, D.F. (1997) Imaging of
oxygen in the surface and deep areas of the kidney. Adv. Exptl. Med. Biol. 411: 591-596.
Phosphorescence O2 Measurement System