REF: DA0650 VOLUME REDUCTION INSTRUMENT Instruction Manual 1.0 Original instructions NOTICE: For safe and proper use, follow these instructions. Please keep them for future reference.
Copyright Any distribution, alteration, translation or reproduction, partial or total, of this document is strictly prohibited unless with prior authorization in writing from Menarini Silicon Biosystems with the exception of the following actions: l Printing all or part of the document in its original format. l Copying contents without any modification and stating Menarini Silicon Biosystems as copyright owner.
Contents Glossary of terms 4 Assistance 4 Information on this manual 5 Safety Warnings 6 6 Reception Unpacking Identification 11 11 12 Product description Introduction to VR NxT Instrument structure Closed holder structure 13 13 14 15 Getting started Installing the instrument Basic operations with the instrument Basic operations with the holder Display description 16 16 17 18 23 Operating with VR NxT Process for general purposes Process for using DEPArray Programs Volume reduction to 13.
Glossary of terms D DABUF DEPArray Buffer for fixed cells P PBS Phosphate Buffered Saline Assistance How to contact Technical Support In case of malfunction or fault or to obtain information contact Menarini Silicon Biosystems at vrnxt@siliconbiosystems.com. Customer Service (for orders only) Please see the Contact us section at www.siliconbiosystems.com.
INFORMATION ON THIS MANUAL Obligations NOTICE: Read this manual before using the instrument the first time. This manual is an integral part of the product and must be kept for its entire working life. It must be consulted in all situations related to the life cycle of the product, from its delivery through to decommissioning. It must be conserved so that it is accessible to operators, in a clean location and in good condition. In the event of manual loss or damage, contact Menarini Silicon Biosystems.
SAFETY Warnings Electrical risks l l l l l l l l Do not pour water or any other liquids over the power supply (risk of electrical short circuits). Ensure the power supply casing is not damaged. Switch off the instrument and unplug the mains cable before cleaning the instrument. Use only the power cord and power supply set provided by the manufacturer. In case the power cord is damaged, switch off the main laboratory circuit breaker and disconnect the cable from the socket and the instrument.
Safety Other risks l l l l l l l l l l l l Handle the instrument and all its components with care, protecting them from mechanical damage. Inspect the main instrument casing and all accessories for damage before each use and replace any damaged accessories. Even minor cracks can lead to serious material damage. The holder is not waterproof. Do not submerge in liquids. Do not use a worn or non-compliant holder.
Safety Risks for the samples l l l l l l l l l l l l l Damaged tubes must not be used. Use only liquids and sample containers indicated in this manual. Buffers and tubes different from those indicated in this manual are not suitable and should not be used. Handle samples and closed holder with samples inside with care to avoid creating bubbles and nullifying the centrifuge effect. Do not process samples with bubbles. In case of bubbles, the output sample volume is not guaranteed.
Safety Safety labels on the instrument Here are the labels that warn about how to avoid risks: Symbol Description See the Instruction manual. Location Risk of electrocution. External power supply shall be kept away from liquid sources (liquid containers, liquid spillages and so on). Intended use VR NxT is a sample preparation instrument intended to be used for the volume reduction of liquids containing or not containing cells. It is for research use only and not for use in diagnostic procedures.
Safety Employer obligations The employer must do what follows: l l l l 10 recruit, train and assign personnel authorized to perform their tasks train assigned personnel and have them follow the safety regulations for each specific task define work procedures ensure that the instruction manual provided by the manufacturer is followed VR NxT | Instruction Manual 1.
RECEPTION Unpacking Precautions on reception On reception of the product, check the integrity of the package and the shock watch on the package itself. In case the package is damaged or tampered with or the shock watch is triggered, contact Technical Support (see "How to contact Technical Support" on page 4). Open the package contents in a laboratory clean environment and store the package for future use. Check the presence of all components and their integrity.
Reception Storage warnings NOTICE: Keep the packing materials to move or store the instrument in future. The instrument must be stored indoor in its original packaging to avoid exposure to adverse weather conditions. Identification Manufacturer's contacts Menarini Silicon Biosystems SpA Via Giuseppe Di Vittorio 21b/3 40013 Castel Maggiore (BO) Italia www.siliconbiosystems.
PRODUCT DESCRIPTION Introduction to VR NxT Description VR NxT is a laboratory instrument that automates the sample volume reduction procedures. It processes up to four samples in parallel and can be used: l l l for liquids containing or not containing cells for both live and fixed cells for general purposes or in association with DEPArray Operating principle The instrument has two volume reduction programs for general purposes and two volume reduction programs for easing the DEPArray use.
Product description Instrument structure Front structure Part A Lid B C D E F G H Name Function l Open and close the rotor chamber. l Protect from moving parts. l Show rotating parts. l House the rotor. Rotor chamber l Protect from moving parts. Closed holder housing House and lock the closed holder. Information LED Signal an anomaly. The anomaly can be solved by the user. l Command the instrument. Display l Indicate the instrument status. l Signals when a problem occurs.
Product description Closed holder structure Structure The closed holder is composed of the following parts: Part A Cap Name Function Not included. Close the holder. Trap the liquid after each run. Cap RFID tag l Identify the cap type. l Guarantee the cap presence and compliance. Holder Contain the tubes. Cavity for the tube cap Align the tube caps. Slot for the tube Contain the tube during the run. Holder clip Fasten the cap to the holder.
GETTING STARTED Installing the instrument Precautions for positioning the instrument Note: The safety of any system incorporating the instrument is the responsibility of the system assembler. l l l l l l l l Use this instrument in a dry and clean environment. The presence of synthetic materials (synthetic clothing, carpets etc.) may cause damaging electrostatic discharges that may lead to erroneous results. Do not use this instrument in close proximity to sources of strong electromagnetic radiation (e.
Getting started Basic operations with the instrument Open/close the rotor chamber Open the rotor chamber: Close the rotor chamber: Inspect the instrument It is important to regularly check the integrity of the instrument and its components. Specifically, before each use check what follows: l l l l If the power supply and the power cord are damaged. If the lid and the casing are damaged and present relevant scratches or cracks (see "Front structure" on page 14).
Getting started Switch on the instrument Switch the instrument on using the main power switch in the rear: the instrument performs a self-diagnostic test. During the test, the rotor performs a partial rotation and then goes back to its correct position, while the instrument checks the internal brake functioning. If no problem or error is detected, the instrument is ready to be used. Otherwise, a message is displayed (see "Errors" on page 37).
Getting started Insert the tubes Close the closed holder VR NxT | Instruction Manual 1.
Getting started Insert the closed holder in the instrument 20 VR NxT | Instruction Manual 1.
Getting started Position the closed holder Open the closed holder VR NxT | Instruction Manual 1.
Getting started Remove the tubes from the holder The use of tweezers should prevent contamination of the samples during tubes handling. Close the removed tubes 22 VR NxT | Instruction Manual 1.
Getting started Display description Available functions The display allows to do what follows: l l Access the menu and perform a few tasks (for example, choose the display language or turn on the chamber lights). Select a processing program. Display structure Part Description A Instrument status B Program name. In order of appearance: C D E 1. Low Volume 2. High Volume 3. DEPArray OUT 4.
Getting started Menu You can only access the menu when no program is running on the instrument. To enter the menu, press Entry Select language Unlock lid Chamber lights Cleaning System info Update Exit 24 and at the same time for about six seconds. Function Select the display language. Manually unlock the lid when a problem occurs (for example power failure). Turn on the chamber lights (for example to verify the absence of foreign bodies or unexpected liquids).
OPERATING WITH VR NxT Process for general purposes Workflow for volume reduction to 13.5 or 12.5 µl (High Volume) The workflow is the following: = manual intervention = swing bucket centrifuge Workflow for volume reduction to 3 or 2 µl (Low Volume) The workflow is the following: = manual intervention = swing bucket centrifuge VR NxT | Instruction Manual 1.
Operating with VR NxT Process for using DEPArray Workflow for volume reduction to 12.5 µl (DEPArray IN) The workflow is the following: = manual intervention = swing bucket centrifuge Workflow for volume reduction to 2 µl (DEPArray OUT) The workflow is the following: = manual intervention = swing bucket centrifuge Programs Available programs The available programs are the following: l l For general purposes: High Volume and Low Volume. For using DEPArray: DEPArray IN and DEPArray OUT.
Operating with VR NxT Please comply with the following table to obtain the volumes indicated in this Instruction Manual. MAXYMum Recovery MicroAmp Reaction Tube Thin Wall PCR Tube with with cap flat cap Program for general purposes for using DEPArray Programs for general purposes: sample characteristics NOTICE: The correct trap of the liquid in the cap is guaranteed only if the input sample volume is respected. NOTICE: The output sample volume is guaranteed only if the sample does not contain bubbles.
Operating with VR NxT Programs for using DEPArray: sample characteristics NOTICE: The correct trap of the liquid in the cap is guaranteed only if the input sample volume is respected. NOTICE: The output sample volume is guaranteed only if the sample does not contain bubbles. The following table shows the characteristics of the samples for each program intended for using DEPArray.
Operating with VR NxT 3. 4. 5. 6. Place the holder rack with the holder under the biosafety cabinet class II. Open the tubes and insert them in the holder one at a time. Start from number 1 position indicated on the holder. Insert each tube cap in its tube cap cavity to align them (see "Insert the tubes" on page 19). Put a new cap on the holder and push the holder clips to close the holder. NOTICE: Do not remove, damage or write on the RFID tag of the cap.
Operating with VR NxT Volume reduction to 3 or 2 µl Warnings l l l Do not lean on the instrument. Do not stay longer than necessary within the 30 cm of clearance from the instrument. Do not deposit any potentially hazardous material within the 30 cm of clearance from the instrument. Notices about the input sample NOTICE: Avoid inserting samples with bubbles in VR NxT. In case of bubbles, the output sample volume is not guaranteed (see "Troubleshooting" on page 37 to solve the problem).
Operating with VR NxT 4. At the end of the program, open the rotor chamber. NOTICE: Do not open the rotor chamber while the program is running and the status LED is steady. 5. 6. 7. 8. 9. Move the closed holder in the holder rack. Close the rotor chamber. Move the holder rack with the closed holder under the biosafety cabinet class II. Push the bottom of the holder clips to open the closed holder. Lift the cap and throw it in the biohazard waste disposal container.
Operating with VR NxT Perform a controlled stop To stop the rotor gently, press . NOTICE: Before opening the rotor chamber, check from the transparent lid if the rotating parts have completely stopped. Perform a quick stop To stop the rotor immediately, choose one of the following solutions: l Turn off the main power switch. Unplug the power supply from the instrument (see "Stop the volume reduction process" on the previous page) pulling the locking device backwards.
Operating with VR NxT Reduce the volume to 13.5 or 12.5 µl if the volume is similar to the starting level 1. Take each tube with the tweezers (see "Remove the tubes from the holder" on page 22), move it to a common tube storage rack and close it (see "Close the removed tubes" on page 22). 2. Move the tubes in the centrifuge racks. 3. Centrifuge the tubes using a swing bucket centrifuge. 4. "Prepare the closed holder" on page 28. 5. "Load VR NxT" on page 29. 6. "Reduce the volume to 13.5 or 12.
MAINTENANCE OF THE INSTRUMENT Cleaning Precautions l l l l l l l Particularly aggressive detergents can damage casing and lid. These detergents can crack the plastic, bleach and/or soften the material. Before using any cleaning or decontamination methods except those recommended by Menarini Silicon Biosystems, check with Menarini Silicon Biosystems itself that the proposed method will not damage the equipment.
Maintenance of the instrument 5. Open the rotor chamber and clean the four rotor holes with the hole brush. Reagents are not required. 6. Close the rotor chamber. 7. To move the rotor back in its correct position, connect the instrument to the power supply and switch it on. Clean the internal parts of the rotor chamber 1. 2. 3. 4. 5. 6. Switch off the instrument and disconnect it from the power supply. Wear gloves. Open the lid. Remove any foreign body from the rotor chamber.
Maintenance of the instrument 3. Use the swab small head to clean the internal part of the holder. 4. Prepare a solution made of water and a 70% Ethanol solution and put it on another swab small head. 5. Use this swab small head to remove any residue of DNA AWAY from the holder internal part. 6. Clean the holes with the hole brush. 7. 8. 9. 10. 11. Clean the holder surface with a damp lint-free cloth. Put the DNA AWAY on the cloth and clean the holder surface again.
TROUBLESHOOTING AND ERRORS Troubleshooting Situation There is a bubble in the sample. The instrument shakes/makes unusual noise when turned on. Not ready is displayed and the status LED is off. A malfunction occurs/something went wrong in the rotor chamber. What to do 1. 2. 1. 2. Centrifuge the sample. Try to aspirate the bubble with a pipette. Stop the instrument (see "Stop the volume reduction process" on page 31).
Troubleshooting and errors Perform a VR NxT diagnostic test 1. Ensure no closed holder is in the closed holder housing. 2. Turn on the instrument: the self-diagnostic test starts automatically. When errors cannot be solved Please contact Technical Support (see "How to contact Technical Support" on page 4). Error codes Below is a list of error codes that can be displayed, together with a description of what might have caused them and a possible solution.
Troubleshooting and errors Code Cause ED028 Rotor not aligned ED029 Closed holder not correctly inserted ED030 Lid lock/unlock failure ED031 At the end of the process, the closed holder housing has not reached the right position. ED032 RFID comunication problem ED033 The samples have not been correctly processed due to an out of range rotor speed during the process. 1. 2. 3. 1. 2. 1. 2. 3. 1. 2. Solution Ensure no foreign body is in the rotor chamber. Turn off and on the instrument.
TECHNICAL FEATURES Technical data Features Throughput (cycle time) l l DEPArray IN and High Volume: 4 samples in ~25 s DEPArray OUT and Low Volume: 4 samples in ~7 s Permitted consumable Permitted holder DA0644 (provided in a box of 72 pcs, REF: DA0665) l Blue holder (DA0649) l Gray holder (DA0648) Max rotational speed 3600 rpm Maximum kinetic energy 10 J Time to stop the instrument l l Quick stop: < 250 ms Controlled stop: < 3 s Sample container 200 µl PCR tube (see "Required materials and too
Technical features Electrical data Power connection See "Power cord" below Power consumption 120 W Mains supply voltage(referred to power supply) 100-240 V AC / 50-60 Hz Mains supply voltage fluctuation Not to exceed ± 10% of nominal voltage Power supply type Mean Well GSM120B24-R7B Power supply output 24 V dc–5 A Input protection 5 A fuse inside the system, not replaceable I/O ports n. 1 USB 2.0 female type A: for software update n.
Technical features Ordering information Consumables Picture Name Description Provider Quantity Code Menarini Silicon Biosystems 72 pcs per cap box Cap box: DA0665 (cap: DA0644) 20 vials per cap box KI0066 Cap Cap to close the holder and trap the removed liquid after each run. DEPArray Buffer for fixed cells Buffer to maximize the Menarini sorting performances of fixed Silicon cell samples.
Technical features Required materials and tools (not included) Image Name Provider 70% Ethanol solution Any supplier MicroAmp Reaction Tube with cap, 0.2 mL, autoclaved Thermo Fisher Scientific MAXYMum Recovery Thin Wall PCR Tube with flat Corning Axygen cap, 0.
Technical features Image Provider Code Thermo Scientific DNA AWAY surface decontaminant Name Thermo Fisher Scientific 7010 Centrifuge rack BIOPlastic (suggested) BP-1 B10420 (suggested) Tweezers Any supplier Generic Disposal Disposal liability As specified by the European Directive 2012/19/EU, never dispose of the product as standard urban waste.
ANNEX A: CELLSEARCH SAMPLE PREPARATION FOR DEPArray Prepare the CELLSEARCH sample for DEPArray with VR NxT Materials required Name Volume Provider Corning Thick Gel Loading Pipet Tip 1-200 μl Round 0.5 mm Corning Protein LoBind tube 1.5 mL Eppendorf Code 4853 l l Research plus pipette 20-200 μl Eppendorf 100-1000 μl epT.I.P.S LoRetention ep Dualfilter 20-300 μl P200 pipette: 3120000054 P1000 pipette: 3120000062 Eppendorf l l 50-1000 μl l l Swing bucket upstream centrifuge for 1.
ANNEX A: CELLSEARCH sample preparation for DEPArray Open the CELLTRACKS cartridge 1. Hold the CELLTRACKS cartridge in one hand. 2. Insert the wide-end of a clean P1000 tip into the opening in the backside of the cartridge, against the top of the insert. 3. Gently push the top edge of the insert to remove the lid of the cartridge. Extract the sample from the CELLTRACKS cartridge 1. 2. 3. 4. 5. 6. Take the P200 pipette with the BSA coated tip and push the pipette knob to the first stop.
ANNEX A: CELLSEARCH sample preparation for DEPArray 7. 8. 9. 10. 11. 12. Take 1,000 μl of DABUF from the DABUF bottle. Put the pipette tip against the wall in the upper part of the Sample tube. Carefully push the knob to release the DABUF and let the liquid flow down the wall of the tube. Discard the tip. Centrifuge the Sample tube at 1,000 g for 5 minutes at room temperature in a swing bucket centrifuge.
ANNEX B: DEPArray RECOVERIES PREPARATION FOR DOWNSTREAM GENETIC ANALYSIS Prepare the DEPArray recoveries for downstream genetic analysis with VR NxT Sample characteristics Note: Samples prepared according to this protocol can be used for downstream analysis methods such as Ampli1 WGA Kit or similar. Cells are collected from the DEPArray following the rules below: Number of cells one up to 20 more than 20 Number of drops* one two six Note*: Number of cell collection drops from DEPArray.
ANNEX B: DEPArray recoveries preparation for downstream genetic analysis 8. If the samples contain two drops, refer to the following table: If samples come And the contained Then remove from the from... cells are... meniscus... DEPArray V2 fixed DEPArray NxT live fixed 20 µl of cell medium - live 16 µl of cell medium 9. If the samples contain six drops, refer to the following table: If samples come And the contained Then remove from the from... cells are... meniscus...
LICENSE AGREEMENT Software conditions of use l l l License Agreement l Menarini Silicon Biosystems ("MSB") licenses the Software installed in VR™ NxT product to you subject to the terms contained in this end user license agreement (this "Agreement" or "EULA"). l l You should read the terms and conditions of this agreement carefully before using VR™ NxT system.
License Agreement (e) reverse engineer, decompile, or disassemble or otherwise attempt to derive the source code; (f) redistribute, encumber, sell, rent, lease, sublicense, use the Software in a timesharing or service bureau arrangement, or otherwise transfer rights on the Software under any circumstances; (g) remove or alter any trademark, logo, copyright or other proprietary notices, legends, symbols or labels in the Software or on the related data storage devices, packaging and associated documentation;
License Agreement tort, warranty or under any statute (including without limitation any trade practice, unfair competition or other statute of similar import) or on any other basis for special, indirect, incidental, multiple, punitive, or consequential damages arising out of the possession or use of, or the inability to use, the Software or the associated documentation, even if MSB is advised in advance of the possibility of such damages, including without limitation damages arising from or related to loss
CONFORMITY Declarations Declaration of conformity Following is the list of Directives the product meets: l l l l 2006/42/EC (MD - Machinery) 2014/30/EU (EMC - Electromagnetic compatibility) 2014/53/EU (RED - Radio equipment) 2011/65/EU (RoHS - Hazardous substances in electrical and electronic equipment Standards applied are: l l l l l l l l l l EN ISO 12100 ISO/TR 14121-2 EN ISO 13849-1 EN ISO 14119 EN ISO 14120 EN 60204-1 EN ISO 7010 EN 61010-1 EN 61010-2-020 IEC 61326-1 FCC statement The System compli
WARRANTY AND RESPONSIBILITY Warranty l Limited warranty l The warranty period begins from the date of receipt of the instrument (if marketed directly by Menarini Silicon Biosystems); consult terms of sale for warranty period. l l l Menarini Silicon Biosystems warrants to the Customer that VR NxT instrument is free of all material and labour defects for the aforementioned warranty period.
Warranty and Responsibility Customer The Customer must inform Menarini Silicon Biosystems of any problems detected as quickly as possible from the moment they occur, describing operations on the device that took place just before and after the breakdown. It must also make any information requested available to technicians during the breakdown analysis process. The device must be available for Menarini Silicon Biosystems maintenance technicians for the time needed to recover it.
Menarini Silicon Biosystems SpA Via Giuseppe Di Vittorio 21b/3 40013 Castel Maggiore (BO) Italia www.siliconbiosystems.com Instruction Manual VRNXTinstr1EN 1.
