AtmosLM Large Molecule Sampling Manual
Contents
1. the 10 uL min flow rate and insert the probe into the animal via guide cannula that has been implanted in advance If dummy probe is hard to remove please twist and pull Maintain the higher flow rate for 2 3 hours in order to prevent clogging of the tubing Then lower the flow rate to 1 uL min or your desired flow rate If necessary refill the syringe of the syringe pump or switch the syringe by using a liquid switching valve such as the Sl 60 Tips Set the flow on the peristaltic pump 5 higher than syringe pump 0021 1 05 ul min for the peristaltic pump when 1 uL min on the syringe speed When you lower the flow speed stopping or lowering the peristaltic pump first and then do the same on the syringe pump to avoid a large air segment in the outlet tube which leads a difficulty of withdrawing sample Clogging mostly happens on the tubing 3 or outlet of the probe after the vent if it happens To fix please disconnect the swivel and replace to a 1 mL syringe containing water at the tubing 3 out see Fig 3 and Fig 4 Then infuse water toward the probe via tubing 3 The flush will come out from the vent and you can use water Fig 4 Removing Clog 13 Maintenance After completing sampling remove the probe from the system and bypass where the probe was Open the Tubing Guide on the ERP 10 to prevent tubing occlusion and excess wear of the tubing Run 50 100 times diluted chlorine bleach in the entire tubing from the2. USER S MANUAL AtmosLM Large Molecule Sampling Manual 1 System Set Up 2 Operation of Peristaltic Pump ERP 10 Figure 1 shows a typical AtmosLM setup A syringe pump is used to infuse One RS 5S Peristaltic Tubing can be installed on each side of the roller perfusate and the peristaltic pump is used to withdraw sample from the upper and lower Tubing Guide is the groove where the peristaltic tubing probe Always keep the peristaltic pump ERP 10 at the same level as or RT 5S is placed To move the tubing into place against the rollers please not more than 50 cm above the AtmosLM probe to avoid reduced flow from pull and slide the Knob until you hear a click The rotation speed shown in the outlet line of the probe the display is NOT the RPM or flow rate but represents an indication of the rotation speed of the roller head When you use the RT 5S tubing and set Name Ss a Specification the speed to 0020 the actual flow rate will be 1 uL min Do NOT place the Model Specification ERP 10 higher than approx 50 cm 20 inches above the probe to avoid Syringe Pump ESP 32 1 motor 2 drive the flow fluctuations In order to prolong the life of the RT 5S please open the Tubing Guides such that they don t compress the tubing whenever the Push Pump ESP 64 2 motor 4 drive ae pump is being stored peristaltic Pump ERP 10 giaa oie Fraction Collector EFC 82 Up Increases the rotation speed of roller EFR 82 Cooling Unit Down Decreases the rotat
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4. ion speed of roller Freely Moving System ATFM RA CW CCW Selects the direction of roller rotation CW indicates ATFM MA Clock Wise and CCW is Counter Clock Wise If held down for 3 seconds the rotation speed will be set to AtmosLM Probe PEPAY Polyethylene 1 000 500 If held for 5 seconds it will be to 1000 kDa Start Stop Starts or Stops the rotation of roller If held down for 5 seconds the input value will be memorized and the same value will return after you reboot the unit Peristaltic Pump Display Fraction Collector Operations Button Tubing Guide Roller Knob Fig 1 An Example of System Setup Fig 2 Peristaltic Pump ERP 10 www eicom usa com info eicom usa com www eicomeurope com info eicomeurope com USER S MANUAL E COM A 3 Tubing Setup 4 Peristaltic Tubing Please always use Eicom RT 5S for the tubing set on the ERP 10 Please exchange RT 5S to new one at least every two 2 weeks if you use every day The lifetime for storage is 6 months When you do not use the ERP 10 please open the tubing guide to relieve the pressure on the RI 5S RT 5S Includes Connect all tubing as shown in Figure 3 Table 1 shows the model names of tubing size material and approximately internal volume of each tube shown in Fig 3 The tubing between the probe and peristaltic pull pump has to be 0 25 mm ID to allow smooth withdrawal of the sample Peristaltic Tubing 5 pcs Joint RTJ 4 pcs Inte
5. mL syringe Larger syringes can easily generate too much pressure Then cover the probe s vent holes with your fingers and depress the syringe plunger gently to fill the probe Water should be coming out from Inlet needle Confirm that there are no leaks from tip of membrane or connecting parts of membrane or shaft The surface of membrane may have sweat which is normal 10 Activate Membrane Submerge the membrane in ethanol 70 to 100 it does not matter for 1 second Do NOT keep the probes in ethanol for longer than 5 seconds as this can damage the probe by weakening the glue The membrane is properly wetted when the opaque white membrane becomes slightly translucent Now use the plastic syringe to push water into the probe again 11 Connecting Probe to System Both the syringe and peristaltic pumps should still be stopped and flow rate settings of the syringe pump is at 10 uL min and ERP 10 is showing 0200 Remove the bypass where the probe will be placed and connect the inlet and outlet Start the syringe pump and start the peristaltic pump immediately after confirming a little leak from the vent holes Pay attention there might be a lag time on the syringe pump caused by a mechanical play see section 6 Confirm that an air segment no larger than approx 5 cm 2 inch 2 5 uL has entered the outlet tubing see 3 4 or 5 on Fig 3 www eicom usa com info eicom usa com USER S MANUAL 12 Insertion of Probe Maintain
6. rnal Diameter 0 25 mm Total Length 83 mm 5 You will need aCSF BSA Bovine Serum Albumin 0 2 um syringe filter one 5 mL plastic syringe one 2 5 mL or 1 mL disposable syringe ethanol Fig 3 Tubing Diagram 6 Flushing System with Water Details of each tubing and parts on figure is explained on Table 1 Please fnd vorrespandinanuiiber inthe fable and eae Flush tubing with water to confirm the flow At this point please bypass AtmosLM probe by using a small connector to join the tubing where the probe will be located If the traveling time of sample is important we recommend checking the actual traveling time on your system setup Insert an air gap in the tubing and run the system Many syringe pumps have a lag time between when the pump is started at the controller and when the actual flow begins This is because there is a play in the cam and motor gear Table 1 Information of Each Tubing Model Name Size Material Approx Volume 1 Tubing from Syringe to Swivel JT 10 50 0 1 mm ID x 50 cm Teflon Tubing Rat WT 35 40 0 1 mm ID x 40 cm FEP 3 1 UL 2 Tubing from Swivel to Probe Mouse WT 25 25 0 1 mm ID x 25 cm FEP 0 6 uL Rat WT 35 40 0 25 mm ID x 40 cm FEP 20 uL 3 Tubing from Probe to Swivel Mouse WT 25 25 0 25 mm ID x 25 cm FEP 12 uL 4 Tubing from Swivel to ERP 10 JF 10 50 0 25 mm ID x 50 cm 2 il JT 10 50 A ID Tefl Auk 5 Tubing from ERP 10 to Fraction i i a ai i Collector Needle S FRN JJ 0 12 mm ID
7. syringe pump to the fraction collector and let it stand for at least 15 min Then disconnect all tubing and flush each piece of tubing with water If you plan on storing the tubing for more than a few days run ethanol and then air through the tubing and store them dry www eicomeurope com info eicomeurope com USER S MANUAL Eico SA Appendix 1 AtmosLM Guide Cannula Implant N Stereotaxic Apparatus Adaptor PESG X Guide Cannula PEG X Dummy Cannula PED X Anchor Screw AN 3 Cap Nut AC 5 1 Set Guide Cannula on Adaptor and Fix with a Cap Nut Cap Nut AC 5 Stero Adapter PESG X Guide Cannula PEG X 2 Clamp the PESG X Please use standard electrode clamp which is provided from your stereotaxic apparatus manufacturer Eicom does not carry this part Model Number Kopf 1770 1771 1773 Stoelting 51631 51632 51634 Clamp of Stereotaxic Apparatus not provided from Eico a 3 Mount Cement Guide cannula crown is made of acrylic resin and fuse with cement Cap Nut does not Please mount dental cement below the thread of the guide cannula Dental Cement Ancor Screw AN 3 Dummy Cannula PED X 4 Replace Adaptor with Dummy Cannula After confirming the cement is dry and firm loose the cap nut and remove the PESG X from the PEG X guide cannula Then put PED X dummy cannula in to the guide and fasten the cap nut www ei
8. x 11 cm PEEK 1 3 uL 6 Liquid Swivel TCS2 21 0 45 mm ID for both in and out Stainless Steel 316 In 11 uL Out 10 uL SN RT 5S Tygon Vinyl chloride 0 25 mm ID x 83 mm 7 Peristaltic Pump Tub d Joint Total 6 uL A EA RTJ Stainless Steel 26G x 15 mm leroy Joint Tubing JB 30 Biton 0 3 mm ID x your cut Customized tube lengths are available for JT 10 JF 10 and WT 35 www eicom usa com info eicom usa com www eicomeurope com info eicomeurope com E COM A So please first fast forward pump and then switch to normal flow rate before connecting the tubing T aCSF 0 15 BSA Prepare aCSF containing 0 15 BSA Bovine Serum Albumin on the day of sample collection Please do not include buffer components such as carbonate or phosphate in aCSF It will have more chance to clog in the line during in Vivo use Filter aCSF with a 0 2 um membrane filter and fill a Gastight Syringe using on a syringe pump 8 Flushing System with aCSF A connector is still being used in place of the probe Run both the syringe pump and peristaltic pump at 10 times the normal sampling flow rate to fill the system To set the peristaltic pump at 10 uL min please enter 0200 to the speed After filling all lines without the probe with aCSF 0 15 BSA stop both pumps at the same time 9 Quality Check of Probe Connect a disposable plastic syringe filled with pure distilled water to the OUTLET shorter needle of the probe Only use a 1 mL or 2 5
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