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APOLLO 4000

World’s first fully-integrated Free Radical Analyzer

INSTRUCTION MANUAL

Serial No._____________________

061506

World Precision Instruments

www

.wpiinc.com

APOLLO 4000

WARNING

This instrument must not be connected to a local

network nor to the Internet. Do not attach any

peripheral device other than a USB printer. Any

change to the proprietary hard disk registry in this

device — whether by virus or by normally benign

hardware or software installations — may render the

drive or the Apollo software inoperable, requiring the

instrument’s return to the factory for reformatting.

File corruption or damage to applications or

operating system caused by such use will not be

covered by the Warranty.

Summary of Contents for APOLLO 4000

Page 1: ...the Internet Do not attach any peripheral device other than a USB printer Any change to the proprietary hard disk registry in this device whether by virus or by normally benign hardware or software in...

Page 2: ......

Page 3: ...CT NITRIC OXIDE C 1 Initial Set up C 1 Calibration of the NO Sensor C 1 The Calibration Kit C 1 Calibration by the chemical generation of NO C 2 Calibration of NO sensor by decomposition of SNAP C 7 C...

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Page 5: ...lated multi channel electrode based free radical analyzer designed specifically for the detection of a variety of redox reactive species of biomedical importance The electrochemical amperometric detec...

Page 6: ...based on a 24 bit A to D and 16 bit D to A conversion driven by a Digital Signal Processor DSP The User Interface is built on a standard PC platform with Windows 2000 operating system A standard seria...

Page 7: ...erformance Faraday shield designed to minimize susceptibility to environmental noise The Apollo software can be programmed to display either redox current e g pA or concentration e g nM Output from th...

Page 8: ...hannels Temperature LED red Sensor LED green Power Temperature Input Sensor Input DVD CD RW Drive Mains Power 110V 220V Switch mouse Power Supply for Monitor Keyboard Analog Outputs BNC USB Ethernet S...

Page 9: ...nd Returns on the Warranty page of this manual Returns Do not return any goods to WPI without obtaining prior approval RMA required and instructions from our Returns Department Goods returned unautho...

Page 10: ...APOLLO 4000 INTRODUCTION WORLD PRECISION INSTRUMENTS A 6...

Page 11: ...ther end of the voltage converter to a matching 3 prong grounded wall receptacle and switch on 4 Connect the power supply cord to the back of the APOLLO 4000 and plug the other end into a matching 3 p...

Page 12: ...by WPI vary in their fragility However at all times the user must exercise caution to avoid damaging the delicate polymeric membrane covering the end of each sensor This membrane prevents water and d...

Page 13: ...g LED indications will inform the user of the status of the system LED CONDITION INDICATION Temp LED red Steady RED light Electrode is performing normally No RED light Electrode is not connected or is...

Page 14: ...re being turned off In the example shown the set up is for a four channel nitric oxide application with horizonal scale of 6 sec division and vertical scales 5000 pA div for channels 1 to 4 Data for e...

Page 15: ...Open opens an existing data file Save saves the current experiment with the default name Save as allows the user to chose the name and the path of the saving data file Print prints the screen to the s...

Page 16: ...le structure and it is therefore necessary to close the previous file Fig B4 This can interrupt the current experiment which may be undesirable Therefore the user must carefully select the sampling ra...

Page 17: ...nA range is selected then the noise floor will be approximately 10 nA divided by 1 000 000 i e 10 fA Conversely if 10 A range is selected then the noise floor will be approximately 10 A divided by 1 0...

Page 18: ...y configures the poise voltage i e 700 mV on the selected channel to measure oxygen Hydrogen Peroxide Automatically configures the poise voltage i e 400 mV on the selected channel to measure hydrogen...

Page 19: ...submenu in the Setup control The Amplitude Unit Conversion screen appears Fig B13 Example The user defines a new unit called Custom with the conversion ratio 1 unit 186 pA Fig B14 If Add Unit button...

Page 20: ...on pushed There are a total of two cursors The second cursor appears after the first one when the pointer device left button is released Calculate Menu includes five different self explanatory paramet...

Page 21: ...set each channel individually all together or unzero all of previously zeroed channels The subtracting value is acquired at the moment of selecting the appropriate action Zero operation can be applied...

Page 22: ...nt the Note to be added see Fig B22 This will create a mark at that place Double click the created mark A Note window will open where a message of up to 64 characters can be typed in see Fig B23 After...

Page 23: ...a real signal application An example file is loaded in the Apollo 4000 Fig B24 Note the temperature sensor is also connected to channel 1 i e 25 C line However the sensor signal is not seen If we appl...

Page 24: ...user must double click inside the appropriate channel plot The second window has a distinctive yellow background color Only one second window can be opened at the same time The size of the second wind...

Page 25: ...sed on the decomposition of the S nitrosothiol NO donor SNAP using either Cu I as described in Method I or Cu II as outlined in Method II as a catalyst The NO liberated from SNAP is used to calibrate...

Page 26: ...ate Standard Nitrite is available from WPI catalog 7357 Alternatively crystalline reagent KNO2 may be used but the user should note that KNO2 is extremely hygroscopic and degrades once exposed to atmo...

Page 27: ...torage 2 Immerse the ISO NOP sensor tip in a strong saline solution 1 M and after waiting a few minutes for the current to stabilize record its value If the current is offscale or unstable after sever...

Page 28: ...has undergone a large temperature change 8 If you feel it is necessary to degas solution 1 prior to calibration this can be done by inserting one of the long stainless steel needles included with the...

Page 29: ...by simple dilution factors as follows 50 L of 50 M KNO2 solution 2 into 10 mL solution 1 1 201 dilution Hence amount of NO produced 50 M 201 0 2487 M 249 nM Likewise Addition 2 100 L of solution 2 add...

Page 30: ...O3 to NO2 Researchers who use the Greiss method for NO2 determination will also be very interested to learn that using the ISO NOP and the titration method for calibration described above a detection...

Page 31: ...rameters involved in SNAP decomposition neither is it intended to propose a model by which SNAP is decomposed Two methods are described here for the calibration of NO sensors based on decomposition of...

Page 32: ...copper II sulfate CuSO4 or cupric II chloride CuCl2 in which Cu II is the active catalyst for the conversion of SNAP to NO Getting Started Prepare the following solutions 1 Saturated solution of cupr...

Page 33: ...commend the use of high grade SNAP with minimal purity of 98 or better SNAP can be purchased from WPI in various amounts Calibration Procedure Within a nitrogen or argon environment place 10 0 mL of s...

Page 34: ...ng Cu II as a catalyst This method of calibration relies on the use of Cu II for the partial but quantifiable conversion of SNAP to NO This procedure can be used as an alternative to the previous meth...

Page 35: ...ains relatively stable for at least 5 hours if kept in refrigerator Note The purity of standard reagent SNAP is very important for the reported data Use high grade SNAP with minimal purity of 95 or be...

Page 36: ...ing the signal output e g in pA vs the concentration of SNAP added at that time Note that every addition of SNAP solution corresponds to a particular NO concentration This will be discussed below Afte...

Page 37: ...f detectable NO will be below the theoretical value because the copper sulfate solution was at equilibrium with ambient air and consequently a portion of the generated NO would have been immediately o...

Page 38: ...0 6 X SNAP Output Current 232 4 nM 139 4 nM 230 pA 462 0 nM 277 2 nM 488 pA 916 8 nM 550 1 nM 1001 pA Y 32 038 1 88X The data from calibration curve indicates that this procedure allows an excellent...

Page 39: ...standards prepared with NO Gas The following method can be used with all NO sensors Preparing an NO Standard This method has the advantage of allowing the user to calibrate NO sensors in the same env...

Page 40: ...ion is used to remove other nitrogen oxides from the NO gas 4 Place 20 mL of distilled preferably deionized water in a small glass vial Seal the vial with a stopper and insert through the stopper a lo...

Page 41: ...nning at maximum capacity with the front panel closed in the down position 8 After the time in step 7 has elapsed turn off the NO source 9 Immediately remove the two needles from the distilled water v...

Page 42: ...should be placed as close as possible to the site of release of NO Durability and Handling The user must excercise caution when handling any NO sensor to avoid actions which could damage the sensor t...

Page 43: ...rces may couple to the system and produce large extraneous signals in the output record The magnitude of this external noise depends on the environment of the laboratory If the interference introduced...

Page 44: ...ng the end of the stainless sleeve The other end of the sleeve is flanged The locking cap is used to attach the sleeve to the probe handle When the sensor is fully assembled i e with locking cap and s...

Page 45: ...or base solution at times both may be necessary for 10 seconds A good indication of a dirty membrane sleeve is a sluggish response or an unusually low sensitivity If these problems are not rectified b...

Page 46: ...p into the handle The electrode should be observed to press gently against the membrane 9 The current displayed on the meter at this time will be high or offscale 10 Suspend the tip of the new assembl...

Page 47: ...ilar device that permits accurate positioning The electrode should be cleaned periodically in distilled water and dried using soft tissue paper Organic contamination can be removed using a mild enzyma...

Page 48: ...APOLLO 4000 NITRIC OXIDE DETECTION WORLD PRECISION INSTRUMENTS C 24...

Page 49: ...f atmospheric pressure parts per million ppm or as a redox current in nanoamperes nA The ISO OXY 2 houses a platinum working electrode and a silver counter reference electrode inside a stainless steel...

Page 50: ...is switched back on again Zero Oxygen Point Calibration After polarization of the ISO OXY 2 as described above a calibration for zero percent oxygen may be carried out in pure nitrogen gas or in wate...

Page 51: ...vely air can be used as the calibration standard but since water vapor does affect the probe reading it is best to use dry air unless the ambient humidity is accurately known Dry air can be obtained b...

Page 52: ...ed using the software At point 3 and point 4 the sensor is exposed to air 21 oxygen and 100 oxygen correspondingly By using the current at 0 0 nM and 100 236 nA a two point calibration curve is built...

Page 53: ...e tip into the water via the top hole Aerate for a few minutes by bubbling air through the side arm of the bottle at a low pressure using a simple aquarium aeration pump The scale reading should be al...

Page 54: ...oxygen concentration when the probe is in water is 8 4 ppm You do not need to correct for the water vapor effect for a ppm calibration since the values in Table 1a are obtained in water saturated air...

Page 55: ...factor to nA or ppm to nA sensitivity of the probe is determined from the slope of the calibration curve and must be typed into the corresponding field see Fig B15 Only after selecting ppm or unit wi...

Page 56: ...r depending on the unit of pressure chosen 1 for atm pt and pH2O are the total pressure of gas and the partial pressures of water respectively pO2 is the partial pressure of oxygen in blood obtained f...

Page 57: ...let water get into the handle The current on the Apollo 4000 should go offscale when the electrode is being rinsed 4 Gently dry the electrode with a soft tissue Kimwipes Be sure to dry thoroughly the...

Page 58: ...Y 2 is not being used it should be stored with the tip suspended in distilled water Clean the Membrane The membrane sleeve itself requires very little maintenance The primary concern is to avoid to da...

Page 59: ...it s recommended that the electrode be disconnected from the instrument and stored with the tip immersed in distilled water This practice will reduce the possibility of a gradual reduction of electro...

Page 60: ...10 0 6 11 8 11 1 10 5 9 8 7 11 5 10 9 10 2 9 6 8 11 2 10 6 10 0 9 4 9 11 0 10 4 9 8 9 2 10 10 7 10 1 9 6 9 0 11 10 5 9 9 9 4 8 8 12 10 3 9 7 9 2 8 6 13 10 1 9 5 9 0 8 5 14 9 9 9 3 8 8 8 3 15 9 7 9 1...

Page 61: ...500 668 0 88 4000 656 0 86 4500 644 0 85 5000 632 0 83 5500 621 0 82 6000 609 0 80 Table 3 Water vapor partial pressure in mm Hg Temp Pv Temp Pv C mm Hg C mm Hg 0 5 20 18 2 5 22 20 4 6 24 22 6 7 26 25...

Page 62: ...Hb g 100 mL C Plasma 5 g 10 g 15 g 20 g 15 0 0302 0 0310 0 0312 0 0316 0 0323 20 0 0277 0 0282 0 0284 0 0287 0 0293 25 0 0257 0 0261 0 0263 0 0265 0 0271 28 0 0246 0 0249 0 0251 0 0253 0 0259 30 0 02...

Page 63: ...00012 which must contain fresh refillable electrolyte WPI 100042 and is separated from the external environment by membrane covering the end of the stainless sleeve The other end of the sleeve is flan...

Page 64: ...chambers WPI NOCHM or NOCHM 4 can be used as an alternative calibration kit specifically for use at different temperature condition Calibration temperatures from 4 40 C can be controlled using an ext...

Page 65: ...he calibration solution 1 After the current is stable zero the baseline 2 Add a known volume of the H2 O2 standard solution to the PBS buffer solution For example add 0 5 10 20 40 80mL H2 O2 standard...

Page 66: ...ate use Clean the Membrane The membrane sleeve itself requires very little maintenance The primary concern is to avoid to damage the membrane and to keep it as clean as possible After each use the mem...

Page 67: ...lue e g 0 20 pA If this occurs then the electrode is in good working order 5 If the electrode is not clean repeat steps 3 and 4 If necessary the ISO NOP Rejuvenator WPI JUV can be used to restore sens...

Page 68: ...h time It may take several hours for the sensor current to reach a low stable value at which time it will be ready for use 12 If the current observed after a few minutes in the saline solution increas...

Page 69: ...ion procedure are correct b The membrane may be heavily contaminated Clean the membrane as described in the section Cleaning the Membrane c The electrolyte inside the sleeve may no longer be functioni...

Page 70: ...APOLLO 4000 TROUBLESHOOTING WORLD PRECISION INSTRUMENTS F 2...

Page 71: ...the probe is immersed into the effluent of a perfusion system or placed directly into the vein of the heart of an animal In this case grounding the external bathing fluid vein or tissue with a stainle...

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Page 73: ...quested All claims for shortage or damage must be made within 10 days after receipt of shipment Claims for lost shipments must be made within 30 days of invoice or other notification of shipment Pleas...

Page 74: ......

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