NanoSight LM14 Operating Manual P551J 2012 NanoSight Ltd. NanoSight Ltd., Minton Park, Amesbury, Wiltshire SP4 7RT, UK 1
Contents IMPORTANT NOTES... 3 USAGE... 3 SAFETY... 3 SERVICING... 4 MAINTENANCE... 4 HANDLING... 4 UNPACKING AND INITIAL INSPECTION... 5 INSTALLATION... 5 RETURNING EQUIPMENT... 6 WARRANTY... 6 TECHNICAL SPECIFICATIONS... 7 LM14 USAGE... 7 CONTROL BOX... 7 ASSEMBLING THE LM14 VIEWING UNIT... 7 INJECTION OF SAMPLE INTO THE LM14 UNIT... 8 LOADING A NEW SAMPLE... 8 CLEANING... 9 STORAGE... 9 TEMPERATURE CONTROL... 10 OPERATION... 10 INITIALISING... 11 TEMPERATURE SETTING... 11 TROUBLESHOOTING AND FAULT FINDING... 12 TECHNICAL SUPPORT AND FEEDBACK... 12 2
NanoSight LM14 Hardware Manual Explanation of Warning Symbols Warning: read instructions to understand possible hazard prior to use. Electric shock hazard Important Notes Usage The NanoSight LM14 device has been designed to be used only in conjunction with and mounted on the optical microscope supplied, and is supplied as an alternative to the standard LM10 viewing unit. The control box is an integral part of the instrument and the LM14 unit cannot be powered separately. The inbuilt temperature controller allows control of the top plate temperature using customized NTA software. Safety The LM14 viewing unit is classified (to BS EN 60825-1 (2001)) as a Class 1 laser device. The instrument contains a Class 3B laser source which must never be removed from the main instrument housing. The plastic screws supplied to hold the top plate in place must be in position and tightened with the attachment tool on the control box power supply prior to turning on the laser power. Laser power should be turned off prior to removing the top-plate screws. The instrument automatically turns the laser off if the top plate is removed while in operation. This should not be overridden by an user On loading the LM14 unit with sample via a syringe, bubbles may occasionally be introduced into the path of the laser beam resulting in a degree of specular reflection. This can be minimised by carefully loading sample in the manner described in this manual. If bubbles are present, they can be visually recognised as bright light scattering centres in the beam path when the unit is being loaded. It is very important that such bubbles be removed, by flushing with further sample, before the beam is viewed via the microscope oculars. Similarly, certain samples may generate bubbles through out-gassing of dissolved gases over extended periods. Before viewing the beam via the microscope oculars, such bubbles must be removed by carefully withdrawing the sample and slowly reintroducing it to allow the bubbles to disperse. If samples repeatedly show evidence of bubble formation, it is advisable to de-gas the sample before analysis. The instrument must not be used in hazardous areas. The instrument is for use in moderate climates only. Never use the equipment in damp or wet conditions. Avoid excessive heat, humidity, dust and vibration. 3
Do not place liquid filled containers on the equipment. Do not use where the equipment may be subjected to dripping or splashing liquids. When using bio or chemically hazardous sample material, it is the responsibility of the operator to determine the requisite protection for each application. Use only the power adapter and other accessories supplied with the instrument. The LM14 system contains no user serviceable part. It should not be modified in any way. Any modification will void the warranty and could make the device unsafe. Use of controls, adjustments or performance of procedures other than those specified herein may result in hazardous radiation exposure. The temperature controlled unit may become hot. Caution should be taken when handling the device. The temperature controller should always be turned off when unattended. Servicing The LM14 system must only be serviced by qualified NanoSight personnel, or approved agents. The LM14 instrument contains no user-serviceable parts and should not be opened by any user. Opening the casing of the LM14 viewing unit or the control box voids all warranties and could expose the user to hazardous voltages or Class 3B laser radiation Maintenance The LM14 system contains no user serviceable parts. It should not be modified in any way. Any modification will void the warranty and could make the device unsafe. The casing of the LM14 unit and the control box should be kept clean with the use of a damp cloth. Do not allow excess moisture to penetrate any part of the system. Do not use solvents. To maintain best functionality and to protect against cross contamination during and following use, the sample cell should be cleaned as described in this manual. The O-rings should be replaced relatively regularly depending on the frequency of use, i.e. monthly for high usage instruments, every 3 to 6 months for lower usage instruments. Handling The NanoSight LM14 incorporates a rugged housing in a sealed case designed to protect the integrity of the instrument. However, it is a sensitive scientific instrument and should be treated as such. The unit contains a static-sensitive laser diode and should never be used in circumstances when a static discharge may damage the diode. During use and when cleaning it is important to ensure that liquid does not enter the inside of the viewing unit. Use only a damp tissue for cleaning, never expose the unit to excess fluids. During sample loading and instrument operation, ensure that the sample chamber is properly sealed and that there is no evidence of any leaking fluids. 4
Between uses of the NanoSight LM14 unit, or for longer term storage, the unit must be cleaned and dried as described in this manual. The top-plate screws should be left completely loose, so that the glass flat is not held under any pressure when not in use. Failure to loosen the top-plate screws between uses can result in damage to the glass flat. Unpacking and Initial Inspection The standard instrument is supplied with the following accessories: NS14C control box Mains power adapter with attached tool USB cable LM14 connector cable Instruction manual Carrying case Spare LM14 viewing chamber seal Inspect the shipping container when the LM10 is received. Carefully check the contents for completeness and condition. The custom-made padded cases supplied should be kept where possible to facilitate shipping should any articles need to be returned at a later date, although replacement cases will be supplied if required. Notify NanoSight Ltd if the contents are incomplete, or if the instrument or accessories appear to be damaged in any way. Keep all damaged packaging, materials and goods for inspection by the carrier. Installation Initial installation will be carried out by qualified NanoSight personnel. In the event that the instrument needs to be relocated the following instructions must be adhered to: Always disconnect the equipment from the mains and ancillary units before moving. The instrument should be located in an area of good ventilation and with sufficient space for safe and efficient operation and maintenance. The supplied control box should be sited to avoid dangers from spillage and splashing. The equipment must be connected to an earthed power supply with a voltage corresponding to that on the power adapter. Ensure that the mains plug is easily accessible to allow the unit to be disconnected from the mains supply. Always use the mains lead supplied. Your sales representative can provide a lead suitable for your country. 5
Returning Equipment If, for any reason, you experience problems with your instrument, contact NanoSight Ltd (support@nanosight.co.uk). In the unlikely event you experience a problem with the LM14 system that requires returning the unit for repair, please contact NanoSight for a P827 Equipment Return form. The P827 must be completed with the following information: Sender s name and address Sender s contact telephone number and email address Complete list of equipment being returned including serial numbers A detailed description of the problem or reason why the equipment is being returned Declaration that if the instrument has been used with bio or chemically hazardous sample material, all equipment has been fully decontaminated before return On receipt of this form NanoSight will provide a Material Returns Number (MRN). An MRN must be obtained from NanoSight before returning any equipment, and should be clearly displayed on the return shipment and included on all subsequent correspondence. Shipping cases will be supplied by NanoSight as required to ensure safe return of the system. Information on packing procedures is provided on the P827 Equipment Return form. Warranty NanoSight warrants that the LM14 system as supplied with its accessories is free from defects in materials and workmanship for a period of one year from shipping to the customer. During this warranty period, NanoSight will, at its discretion, repair or replace defective products. Any liability under this warranty extends only to the replacement value of the equipment. This warranty is void if: The LM14 or its accessories have been partly or completely disassembled, modified or repaired by persons not authorised by NanoSight Ltd. The instrument or instrument system is installed or operated other than in accordance with these operating instructions. No other warranty is expressed or implied. NanoSight is not liable for consequential damages except as limited by English law. 6
Technical specifications LM14 module Size: Weight: Available temperature control: Temperature Accuracy: Time to temperature: Humidity: Casing: Wetted Parts: 140mm x 74mm x 68mm 650g from 8 o C below ambient up to 50 o C +/-1 C 3 minutes (to indication of within 1 C) 5-95% non-condensing Anodised aluminium alloy Anodised stainless steel, optical glass, nylon and viton rubber Control box Size: 170mm 106mm x 62mm Weight: 600g Power supply: 12V, 5.0A Laser Classification Embedded laser: 635nm CW, max power < 50mW 532nm CW, max power < 60mW 405nm CW, max power < 70mW LM14 Usage Controller Box Connect the controller box as follows: 1. Connect the lead for the LM14 sample unit to the black 15-pin socket. 2. Connect the USB temperature lead to the USB sockets on the control box and the PC. 3. Connect the mains power adapter. 4. If required, connect the laser synchronisation cable, needed to control laser pulsing when using the EMCCD camera. 5. The rocker switch on the front of the control box turns the laser power on and off. The temperature readout and control is permanently on whenever the control box is powered. Assembling the LM14 Viewing Unit The LM14 viewing unit can be assembled as follows: 1) Switch off unit. 2) Clean all necessary parts (see below). 3) Lay top plate on glass element and adjust gently until plate locates. 4) Drop in four screws and finger tighten loosely alternately in opposite diagonal pairs. The LM14 unit is now ready to inject a sample. 7
Injection of sample into the LM14 unit Before injecting a sample, the laser MUST be switched off. Failure to take this precautionary step may expose the user to direct Class 3B radiation. Load the sample into the sample chamber using a syringe (without needle). The Luer fittings are designed to accept standard syringe bodies of all sizes. Note that high pressures can inadvertently be generated when large bore (e.g. 5-10 ml) syringe bodies are used. To avoid generating pressures which might result in the sample bypassing the seals in the sample chamber or damaging the window, care must be taken to introduce the sample slowly. Similarly, care should be taken to avoid the introduction of bubbles at this stage. The LM14 sample unit should be tilted such that the syringe in the inlet port is at a lower level than the outlet port, and liquid introduced as slowly as possible, allowing the chamber to be filled against gravity and bubbles to escape from the outlet port. It is recommended that the sample be loaded into the luer port nearest the connector, so that the outlet port is away from the power connector and that a tissue be held lightly at the outlet port to soak up any excess fluid. Occasionally a bubble may be present in the chamber and in the path of the laser beam. This might cause some degree of specular reflection of the laser beam off the bubble surface. The laser must be turned off and the bubble removed before viewing through the microscope. Certain samples may generate bubbles through out-gassing of dissolved gases over extended periods. In both cases, the image collected by the CCD camera supplied will show a high intensity region (or may be completely saturated) indicative of the presence of such bubbles. Before viewing the beam via the microscope oculars, such bubbles must be removed by carefully withdrawing the sample and slowly reintroducing it to allow the bubbles to disperse. If samples repeatedly show evidence of bubble formation, it is advisable to de-gas the sample before analysis. A properly prepared and loaded sample will appear as a clear sample through which the laser beam can be seen as a thin line passing through the sample chamber. If the beam appears very bright and appears to bloom within the sample, the sample is too concentrated and must be further diluted before viewing via the microscope. Loading a New Sample Before extracting a sample, the laser MUST be switched off. Failure to take this precautionary step may expose the user to direct Class 3B radiation. To remove a sample, simply extract using a syringe. The chamber can be rinsed/cleaned by flushing through the chamber using fresh particle-free water or solvent prior to loading with a different sample. In this instance it is recommended that a tissue be held lightly at the outlet port to soak up the excess solvent from flushing. Between samples which differ significantly in terms of solvent, particle type or particle loading, it is advisable to dissemble the top plate and rinse and dry thoroughly the optical element and top plate as described below. 8
Cleaning Before cleaning the system, the laser MUST be switched off. Failure to take this precautionary step may expose the user to direct Class 3B radiation. Depending on sample type and length of use, it may prove necessary to clean the chamber and optical surface of the LM14 sample unit. Once the laser power has been turned off, the unit may be cleaned as follows: 1. Empty the sample chamber by extracting all residual fluid by syringe via the Luer ports. 2. Unscrew the plastic screws securing the top plate (housing the viewing window) and lift off the top plate. 3. Clean the inside of the window by gently wiping it with a tissue wet with either ethanol, acetone or other solvent required to remove all traces of the sample being analysed. 4. Rinse with deionised water. 5. Dry surfaces of the window with optical grade lens cleaning tissue. 6. An air stream (e.g. from a can of compressed air) should be used to blow any residual liquid from the inlet and outlet channels (luer ports) through which the sample is introduced into the chamber. Redry the surfaces with a tissue if necessary. 7. Carefully wipe the optical flat with a lens tissue or cotton bud damped with solvent. 8. When all optical surfaces have been cleaned and are dry, reassemble using only finger-tight pressure to clamp down the top plate using the plastic screws. 9. DO NOT OVERTIGHTEN. Under no circumstances must liquid be allowed to enter the housing of the LM14 unit at any time. This will cause irreparable damage to the laser mounted within the unit. Take care not to abrade or scratch the optical flat surface, or introduce particulates or contaminants onto the surface. If necessary, other cleaning agents may be employed such as dilute mild detergents, or even dilute acid washes prior to a final solvent cleaning step. The NanoSight device is supplied with optical flats which are relatively robust. However, treat all optical surfaces with the same care as would be employed with equivalent surfaces on your microscope. The optical flats are an integral part of the instrument and may need replacing if scratched or damaged. Replacement optical flats can be supplied by NanoSight (pricing on request). If in doubt about the choice of solvent and its compatibility with any part of the NanoSight device, please contact us for further information. Storage Between uses of the NanoSight LM14 unit, or for longer term storage, the unit must be cleaned and dried as described above. The top-plate screws should be left completely loose, so that the glass flat is not held under any pressure for extended time periods. Failure to loosen the top-plate screws between uses can result in damage to the glass flat. 9
Temperature Control Peltier elements within the LM14 operate as a heat pump transferring heat from one side of the peltier to the other (i.e. they push heat from one face to the other making one side hot, the other cold). They are electrically powered and the power can be controlled by precisely varying the voltage (and therefore current) through the peltier elements. There is a thermistor to measure the temperature positioned within the LM14 body. The temperature measurement is fed back to the controller card and the power adjusted to attain the desired temperature. Available temperature control: Temperature Accuracy: Time to temperature: from 8 o C below ambient up to 50 o C +/-1 C 3 minutes (to indication of within 1 C) Before using the temperature controller it is imperative that all warnings associated with its operation, as described below, are read and understood. The temperature controller may become hot. Caution should be taken when handling the device. The temperature controller should always be turned off when unattended. Operation It is imperative that all users have read and understood these concerns: 1. The temperature controller should be turned off if left unattended. 2. The main body of the LM14 laser module should be monitored to ensure this does not overheat. This is possible if the temperature control is set to a high temperature (>40degC) or a low temperature (<ambient) for a long period of time. 3. It is important that the heating is not able to get out of control. This can happen if a high temperature or low temperature is set for a long time. In either case the LM14 will heat up significantly. Should this happen you should turn the temperature controller off in NTA. Alternatively in an extreme case the power jack to the control box can be removed. Note: if this occurs it may not be sufficient to set the temperature lower, the temperature controller should be switched OFF. 4. When cooling the sample to below ambient temperature, the temperature control should only be employed for a maximum of 15 minutes at a time, before being switched off for at least 15 minutes. Prolonged cooling can lead to an excessive build up of heat around the laser module which can cause fluctuations in the laser power and possible long-term damage. Before using the temperature controller it is imperative that all the warnings mentioned above are read and understood. 10
Initialising 1. Switch on power to the LM14 via the control box. 2. Switch on the PC. 3. Allow the PC one minute from logging on for all drivers to be recognized. 4. Run the NTA Temperature Comms program from the icon on the desktop. 5. Load the NanoSight NTA software 6. Select capture after several seconds the temperature sensor box should read the temperature of the LM14 body and the temperature control module will be initialized. Temperature setting 1. When the temperature sensor is initialized, click the On/Off button. 2. Default set temperature is 23degC. To change this temperature, click the box to the right of the temperature sensor and type in the required temperature e.g. click on the small 23.0 in the image above. Type in the temperature you require and click OK. 3. Click the Set button below this. 4. The temperature controller will now work to set the temperature of the top plate to the target temperature. Some overshoot should be expected. Dependent on the accuracy of temperature required up to 30 minutes should be allowed for the sample temperature to fully stabilize. The angled line shows a trace of the temperature over the last 200 seconds. This line is autoscaling but has no labels. 5. A new temperature can be programmed to change the temperature (by repeating steps 2 and 3). 6. When finished with the system press the off button in NTA to turn off the temperature controller. The temperature readout will continue. 11
Troubleshooting and Fault finding If there is no response from the temperature readout, no readout from the temperature control, or either of these readouts freeze, proceed with the following steps: 1. Check that the USB lead is connected to the PC and to the LM14 controller box 2. Restart the PC. 3. Start NTA--> Capture. If no temperature is showing then close NTA. 4. Remove the USB lead from the PC and put it into a different USB slot. 5. Restart NTA --> Capture. If no temperature is showing then close NTA. 6. Remove from the socket the power lead to the LM14 control box. Replace after 10 seconds. 7. Repeat steps 4 to 6 If there is still no temperature readout, the port numbers may have changed and need to be reassigned on the PC. To fix this problem, follow these steps: 1. Right click on My Computer and select Properties. 2. Under the Hardware tab, click on Device Manager. 3. Expand the Ports (COM & LPT) selection. 4. Note the port number assigned to USB Serial Port. 5. In My Documents, open the file nano.ini. Under the [Preferences] section, there is a TemperatureCom= command. This should be set to the number of the port assigned to the USB Serial temperature controller in Device Manager. Technical Support and Feedback If, for any reason, you experience problems with your instrument, contact NanoSight Ltd (support@nanosight.co.uk) or by phone on + 44 (0) 1980 676 060. 12