Product Information Operation Manual Easy Wash 2000

1. Product Information Operation Manual Easy Wash 2000

2. Content1. General Description 32. Requirements 33. Installation 3 3.1. Connections 3 – 5 3.2. Supplement pump 5 3.3. Programming of «EASY WASH 2000» 6 3.4. Verification of water quantity and pressure 7 3.5. Verification of supplement dosage 8 3.6. Recommendation for ELISA Plate Washing 9 4. Regularity of Fountains 9 5. Maintenance 10 5.1. Daily maintenance 10 5.2. Weekly maintenance 10 5.3. 6-monthly maintenance 106. Warranty and Liability 117. Appendices 11 7.1. Technical Specifications 11 7.2. Chemical Resistance 11 7.3. Supplement Composition 12 7.4. Packing List 12 8. Ordering Information 12 Easy Wash 2000

3. 1. General Description: The «EASY WASH 2000» microtiter plate washer has been specially developed for plant-diagnostic application and provides an efficient and uniform washing process, even for sticky plant extracts. Thanks to a special design where the plates are inserted upside down and rinsed from below, clogging of the nozzles with plant material, as often observed with conventional washers, does not occur with «EASY WASH 2000». Up to 100 plates can be washed per hour. «EASY WASH 2000» is operated with tap water to which a defined amount of supplement such as concentrated PBS-Tween is admixed with an adjustable metering pump. With its compact dimensions, this equipment fits in every laboratory. 2. Requirements: Power supply of 100 V - 240 V AC, 50-60 Hz Tapwater with > 1 - 2 bar constant pressure and of drinking water quality, which is usually between 10 and 25 °C. Extreme conditions of physical and chemical properties of tap water, such as pH and conductivity, are buffered and neutralized by using PBST as supplement (see page 7). Use a prefilter (< 100 µm) if tapwater contains considerable amounts of loam, rust or other impurities (see page 8, Art. No. 600070, optional prefilter of 8µm). An ion exchange device is recommended in case of high lime stone content. Deposit of lime stone may lead to blocking of the internal valve after some time. 3. Installation: 3.1. Connections Install «EASY WASH 2000» close to a sink. Make sure that the drain is elevated relative to the sink, otherwise place the washer on a stable box. Connect tap water to the washer and make sure the water tap is completely open and that the connections do not leak. Tap water connection Easy Wash 2000

4. If the tapwater contains considerable amount of particulate matter, insert the optional prefilter. Prefilter Outlet Mount the grey tubing on the outlet and place the other end in the drain. Grey tubing for waste water Attach tubing from the supplement bottle (PBST 25 x) to the connection at the peristaltic pump. The connection is a quick-lock system: just insert tubing until you hear a «click». To release, press the ring of the lock where the tubing is inserted and pull out the tubing. Connection to the peristaltic pump Easy Wash 2000

5. Connect the plugs of the power supply cable of the Easy Wash and of the AC adaptor, then connect to mains. Power supply cable of the Easy Wash AC adaptor Mains 3.2. Supplement pump The supplement pump is already mounted at the rear side of the EasyWash. However, the pump head and tubing can be easily changed when required in function of usage. First unlock the head by pressing the two clamps at both sides of the head and pull the head away from the motor axis. Then open the housing by releasing the two small hooks. The white rollers need to be cleaned from time to time and the tubing replaced. b a a b Remove the cover of the pump head while releasing the small hooks (b). Press the two clamps (a) locking the pump head and pull it away. Easy Wash 2000

6. 3.3. Programming of «EASY WASH 2000»: • Navigate with the «up» and «down» keys through the four different menus. • Timer: Press «ok» then set wash time in seconds with the «up» and «down» keys. Quit with the «ok» key. • Rinse: To rinse the washing head with tap water while pressing «ok». The metering pump is switched off in this modus. Adjust the water pressure: turn the knob «Water Pressure» to regulate the height of fountains to approximately 2-5 cm above the nozzles. Verify that the water volume is 500 - 600 ml per 10 sec otherwise adjust the volume until the required amount is delivered. • Supplement: Set the speed and dosage of the peristaltic metering pump (range: 50- 210 ml/min). To change the settings, turn the knob «Metering Pump» (See Fig. 3, page 7: Calibration curve of the supplement). By pressing «ok», the metering pump will operate alone in this modus (useful for filling the tubing of the supplement). When using PBST 25x as supplement, set the speed at 42 % which corresponds to a dosage of 20 ml supplement per 10 sec (see fig. 3). This results in 1x working strength of PBST for a volume of 500 ml water per 10 sec. • Wash: Washing process (water + supplement) is started by pressing the «Wash» key. All parameters are listed in this window: Settings of water pressure (in %), supplement (in %), and of the timer (in sec). During the wash process the timer is counting down. This function may also be triggered with an optional footswitch connected to the socket at the rear side of the «Easy Wash 2000». • Note: The optional footswitch as well as the «wash» key are only functional in the wash (4) modus. The functions of the other menus (1-3) are operated with the «ok», «up» and «down» keys. Keys and knobs for adjusting washing parameters 1) 2) 3) 4) Easy Wash 2000

7. 3.4. Verification of water quantity and pressure Best washing conditions are achieved with a water volume of approx. 500 ml / 10 sec giving a fountain height of 2-5 cm at a pressure of 0.5 – 1.0 bar. To verify the settings, place the grey outlet tubing in a graduated cylinder and wash for 10 sec. It might be necessary to elevate the washer in order that the complete amount of water can be collected in the cylinder in the sink. If the amount of water is not equal to 500 ml +/- 50 ml, adjust by turning the knob “Water pressure”. If this quantity of water cannot be achieved anymore by adjustment with the knob «Water pressure», please check the manometer on the rear to make sure that the water supply is at least 0.5 bar. If the problem remains, it is possible that the backflow preventers are blocked and need to be cleaned or replaced. Please contact BIOREBA in that case. Graduated cylinder for measuring water quantity Height of fountains during washing Manometer indicating the actual pressure during operation Easy Wash 2000

8. 3.5. Verification of supplement dosage Best washing conditions are achieved with a quantity of 20 ml of PBST 25x that is admixed to 500 ml of water in 10 sec. To verify the settings, take the black tubing out of the bottle containing PBST 25x and place it in a graduated cylinder containing 100 ml of water. Wash for 10 sec, then take the tubing out of the liquid and read the difference to 100 ml. If this amount is less or more than 20 ml +/- 2 ml, adjust by turning the knob “Metering pump”. If this quantity of supplement cannot be achieved anymore by adjustment with the knob «Metering pump», please remove the pump housing at the rear and replace the tubing of the pump. If you need technical assistance, please contact BIOREBA . Cylinder filled with 100 ml of water Remaining water after 10 sec operation Easy Wash 2000

9. 3.6. Recommendation for ELISA plate washing The ideal washing time is 5 seconds. This provides a wash volume of about five times the volume of a 96-well-microtiter plate per run, if the flow rate (water pressure) has been set at approx. 500 ml per 10 sec (see above 3.3. Verification of water quantity and pressure). It is strongly recommended to wash each plate at least twice for 5 sec in two different positions; i.e. the first 5 sec with the well A1 oriented to the back right corner and the second 5 sec with the well A1 turned to the front left corner. After incubation of sticky plant extracts, washing three times 5 sec is recommended. 180° Wash the first 5 sec with well A1 oriented to the back right corner Turn the plate and wash for the second 5 sec with well A1 oriented to the front left corner 4. Regularity of fountains: Not all of the fountains have constantly the same height during operation. This is due to turbulences of the water flow under the nozzles of the washing head above a certain flow rate, which cannot be prevented by any mean. This phenomena, however, has no effect on washing performance, and it can be observed that the fountain height of neighboured nozzles mutually change with a periodicity depending on flow rate. Nevertheless, it is highly recommended to wash each plate twice and to turn it between the two washing cycles (see above) and to check the fountain of each nozzle on a regular base. There is always the risk that nozzles are completely or partially blocked with fine particles. This can be prevented by cleaning the washing head regularly (see below in the next paragraph «Maintenance») to avoid that particles, depositions, and micro-organisms are accumulating in the washing head. Easy Wash 2000

10. 5. Maintenance: Maintenance is necessary to assure a long lifetime and to prevent malfunctioning. 5.1. Daily maintenance : Clean the system with tap water to remove the wash buffer with the menu «RINSE» (see page 4). Spills of buffers should be removed immediately or latest at the end of a working day with a wet cloth. Attention: do not change accidentally the settings of the “Water pressure” and “Metering pump” when cleaning the housing. If the washer is not used for more than two days perform the weekly maintenance! 5.2. Weeklymaintenance: Unscrew the lid of the buffer bottle (close the bottle with a spare lid) and put the end of the tube in a container with distilled water. Press 6-8 times «WASH» in order to rinse the entire system with water. Disassemble the washing head, clean it with a mild laboratory cleaner. Check the nozzles and clean them with the enclosed needle if necessary. Rinse with water and leave the washing head disassembled until the washer is used again. Weekly maintenance: disassemble the washing head and deblock the nozzles with the enclosed needle if necessary 5.3. Six-monthly maintenance: Check the washing head and decalcify if necessary with a weak acid (1-5% w/v acetic or citric acid). Verify again the quantity of water and supplement according to 3.3. and 3.4., see pages 7+8). Disassemble the pump head of the peristaltic pump by pressing the two clamps and pulling it from the driving shaft. If necessary, replace the tubing and reassemble (see page 5). Tubing and pump head are replacement parts (see page 12). Clean the coarse filter which is built in the pressure reducing valve in regular intervals according to purity of your water supply but at least every 6 months. If the tap water contains considerable amounts of loam, rust or other impurities, it is highly recommended to use an additional pre-filter (8 µm). A pre-filter can be ordered separately (see «Ordering Information», page 12). Easy Wash 2000

11. 6. Warranty and Liability BIOREBA AG guarantees that the delivered product has been thoroughly tested and its properties specified herein are fulfilled. No further warranties are given. BIOREBA AG is unable to accept liability for any damage which results from a non-appropriate use of this product (see also our product information „PI_Warranty for BIOREBA Equipment“ on http://www.bioreba.com). 7. Appendices 7.1. Technical Specifications: External power supply: 100 V - 240 V AC, 50/60 Hz. Internal voltage: 12 V DC Power consumption: 30 VA. Operating temperature range: 5 – 40°C Tap water regulation: A) External pressure reducing valve, adjustable with a hexagon pin key, to reduce tap water pressure (2-6 bar constant pressure required) to 1 – 2 bar. External valve includes a coarse filter for the retention of solid matter (> 1mm), which may cause blocking of wash nozzles. B) internal valve electronically adjustable in the range of 0 - 0.5 bar. Supplement pump: Peristaltic pump, adjustable in the range of 50 – 210 ml/min. External control: socket for plug-in for connecting a foot-switch (operates the «Wash» key function). Dimensions (W x D x H): 220 mm x 450 mm x 230 mm Weight: 6.6 kg 7.2. Chemical Resistance: Materials that come in contact with water are made of brass, stainless steel, PVC (polyvinylchloride), and POM (polyoxymethylene). The supplement comes in contact with polypropylene, stainless steel, PVC, and POM. The device has been designed for the use with tap-water in combination with a supplement such as concentrate of phosphate-buffered saline (PBS) containing detergents (Tween 20, Triton X-100 or similar). Avoid use of strong acids and bases, strong or halogenated solvents, and strong oxidizing or reducing agents, which may cause corrosion or decomposition of the device or parts of it. For decalcifying, use a weak acid such as citric or acetic acid at 1‑5 % (w/v) or a commercial decalcifyer for use in household (e.g. for coffee machines) such as Durgol. Easy Wash 2000

12. 7.3. Supplement composition Depending on application, a variety of different washing solutions may be possible. In ELISA tests, one of the most common buffers is PBST because of its cheap ingredients and good performance in terms of neutralizing detrimental effect of pH and high lime stone content of tap water. The composition for the preparation of PBST 25x is given below. At room temperature, this stock solution remains stable, i.e without cristallisation of any ingredients that may be the case for higher concentrated solutions. Composition of PBST 25x: PBST 25 x Ingredients [PBST 1x] 1000 ml 5000 ml 10000 ml NaCl 137 mM 200.0 g 1000.0 g 2.000 kg KH2PO4 1.5 mM 5.0 g 25.0 g 0.050 kg Na2HPO4 8.1 mM 28.8 g 144.0 g 0.288 kg KCl 2.7 mM 5.0 g 25.0 g 0.050 kg Tween 20 0.05 % 12.5 g 62.5 g 0.125 kg 7.5. Packing List (Art. No. 600000 EASY WASH 2000 complete) 1. 1 EASY WASH 2000 2. 1 AC adapter (100 V - 240 V) with plug set 3. 1 Tube with metal jacket for connection to the tap water supply (3/4” connections) 4. 1 Tube (grey) for connection of the drain to the sink 5. 1 glass bottle 1000 ml with modified cap (45 mm) for the supplement 6. 1 Tube (80 cm) for connection of supplement to washer 7. 1 Tube (23 cm) for supplement for use with 1000 ml glass bottle 8. 1 Tube (35 cm) for supplement for use with 5000 ml glass bottle 9. 1 Set of four 1 mm spacers for adjusting height of microtiter plates 10. 1 Needle for unblocking the nozzles of the washing head 11. 1 Replacement set of tubings for metering pump, incl. one pump head 12. 1 Manual 8. Ordering Information Art. No. Description 600000 EASY WASH 2000, 96-well ELISA plate washer complete 600010 Optional foot-switch 600020 Replacement washing head for 96-well plate 600030 Replacement set of tubings for metering pump, incl. one pump head 600040 Replacement cap with connections for glass bottles 600050 Replacement glass bottle 1000 ml 600060 Optional glass bottle 5000 ml 600070 Optional pre-filter housing and filter (8 µm) 600080 Replacement filter inserts (8 µm) for Art. No. 600070 110154 Washing buffer (PBST) powder (1250g) for preparation of 5 litre PBST 25x Easy Wash 2000