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SMARTer™ ICELL8® cx Single-Cell System User Manual

(061918)

takarabio.com

Takara Bio USA, Inc.

Page 12 of 35

Syringe Pump Unit

The Syringe Pumps Unit is a hydro-pneumatic system that controls the aspiration and dispensing of samples and
reagents in the ICELL8 cx instrument. A tubular harness connects the cx Stage Module and the Syringe Pump
Unit. The Syringe Pumps Unit and solenoid valves are used to closely control liquid dispensing.

Figure 4. Syringe Pumps Unit.

cx Stage Module

The cx Stage Module houses the head, tips, cx Dispensing Platform, Multi-Well Plate Nest, Wash Station, Tip
Mount, and XY Transitional axes. The cx Dispensing Platform aspirates reagents from the 365-well plate and
dispenses into a chip. A single interface cable facilitates mechanical control between the two modules. An
environmental chamber surrounds the cx Stage Module to maintain optimal humidity levels during reagent
dispensing.

Figure 5. ICELL8 cx Stage Module.

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Содержание SMARTer ICELL8 cx

Страница 1: ...echnical Support techUS takarabio com United States Canada 800 662 2566 Asia Pacific 1 650 919 7300 Europe 33 0 1 3904 6880 Japan 81 0 77 565 6999 Page 1 of 35 Takara Bio USA Inc SMARTer ICELL8 cx Single Cell System User Manual Cat Nos 640188 640189 640187 640002 640198 061918 ...

Страница 2: ...col Quick Reference Guide 16 VI Protocol Preparing the ICELL8 cx System 17 A Powering on the System 17 B Checking System Containers 17 C Running the Daily Warmup 18 D Running the Tip Clean Procedure 19 VII Protocol Preparing the Source Plates 20 VIII Protocol Dispensing Samples into the Chip 20 A Selecting the SMARTer ICELL8 cx Application 20 B Placing the Chip in the ICELL8 cx Unit 21 C Placing t...

Страница 3: ... source plate 21 Figure 16 View of the ICELL8 cx with the front door closed and open 22 Figure 17 Select the 3 DE TCR tab 22 Figure 18 Vacuum status confirmation 23 Figure 19 Confirm plate and chip loading 23 Figure 20 Confirm seal removal 23 Figure 21 Live image of chip scanning 24 Figure 22 Input chip ID 24 Figure 23 Vacuum status after dispense 24 Figure 24 Initiate chip scanning 25 Figure 25 R...

Страница 4: ...Table III ICELL8 cx Installation Kit components 14 Table IV Materials from TBUSA required for all applications 15 Table V Quick reference guide 16 Table VI Problem Drop dispersion or improper dispensing 34 Table VIII Problem Dispensing head does not home 34 Table IX Problem Low or partial dispenses 34 Table X Problem Apparent low sample concentration 35 Table XI Problem System stalls because the s...

Страница 5: ...or damage to or destruction of the system CAUTION Indicates a hazard that could result in loss of data or damage to the system Indicates the presence of an electrical shock hazard Proceed with caution This symbol may appear next to either a WARNING or CAUTION symbol Indicates the presence of a biological hazard Proceed with caution This symbol may appear next to either a WARNING or CAUTION symbol ...

Страница 6: ...personal protective equipment PPE which should at minimum include gloves eye protection and a lab coat However the choice of PPE used should be dictated by the biosafety level of the biological samples being introduced into the ICELL8 cx system Please consult your institutional biosafety committee for additional information on the necessary precautions for your sample type WARNING Class I Equipmen...

Страница 7: ...rument is designed to withstand safely It depends on the nature of the electricity supply and its means of overvoltage protection For example in CAT II which is the category typically used for instruments in hospital research and industrial laboratories the expected transient overvoltage is 2 500 V for a 230 V supply and 1 500 V for a 120 V supply Pollution Degree 2 according to IEC 60664 1 Pollut...

Страница 8: ...ving the system WARNING If you need to move the system after it has been installed please contact Takara Biosystems Inc Warning labels on the instrument Please note the warning label on the instrument WARNING This system contains moving parts Keep hands away from the system while the instrument is in use ...

Страница 9: ...tion Kit 1 1 x ICELL8 Chip Alignment Film 2 x 384 Well Source Plate Seal 1 x ICELL8 Alignment Chip 1 x Imitation Master Mix with UV Dye Rox 45 ml 1 x Blotting Paper 2 x 384 Well Source Plate Regional Power Cord 5 SMARTer ICELL8 cx Loading Kit 3 2 x Blotting Paper 1 x Optical Imaging Film 1 x TE Sealing Film 1 x SmartChip Intermediate Film SMARTer ICELL8 Collection Kit 3 1 x Collection Fixture 2 x ...

Страница 10: ...Reservoir and Electronic Scale Wash Container Humidifier Peristaltic pump Desktop and Monitor Fluidic Harness and Power Cord Waste Container SMARTer ICELL8 cx Software User Manual this document Tool Set Blotter and Balance Plate Water Container and Electronic Scale The Water Bottle contains deionized Milli Q water or equivalent that occupies all fluid paths in the ICELL8 cx instrument The liquid i...

Страница 11: ... Figure 2 Water Bottle seated on the Electronic Scale Humidifier The Humidifier maintains the relative humidity at 45 in the Environmental Chamber to minimize reagent evaporation during the dispensing process The Humidifier is behind the back cover of the ICELL8 cx unit Figure 3 Figure 3 Humidifier unit ...

Страница 12: ...nd solenoid valves are used to closely control liquid dispensing Figure 4 Syringe Pumps Unit cx Stage Module The cx Stage Module houses the head tips cx Dispensing Platform Multi Well Plate Nest Wash Station Tip Mount and XY Transitional axes The cx Dispensing Platform aspirates reagents from the 365 well plate and dispenses into a chip A single interface cable facilitates mechanical control betwe...

Страница 13: ... Category Specification Dispense volume 50 nl or 100 nl per nanowell Software SMARTer ICELL8 cx Software Laptop computer Windows 7 2 GB memory 120 GB storage 1 GB network adapter USB ports for memory sticks Power requirements for different electrical grid types 120 VAC 60 Hz mains four 120 V 15 or 20A circuits 220 240 VAC 50 Hz mains four 10A outlets 100 VAC 50 60 Hz mains four 15A outlets Fuses D...

Страница 14: ...n s IT personnel to avoid interfering with instrument operation NOTE To avoid contaminating your PCR do not install the ICELL8 cx in an area that could contain high copy DNA or amplicons from previous PCR experiments C Required Equipment and Supplies from Your Lab or Other Suppliers Wash Solution 500 ml of 0 2 sodium hypochlorite solution made from reagent grade sodium hypochlorite in deionized fi...

Страница 15: ...specific 384 well plates are the required container for solutions that will be dispensed using the ICELL8 cx Plate seals are included IV System Description SmartChip Technology SmartChip technology distinguishes TBUSA s platform from other systems Each chip has a 72 x 72 array of nanowells and can accommodate up to 5 184 100 nl real time PCRs in a single run The ICELL8 cx unit transfers samples co...

Страница 16: ...he Tip Clean procedure 2 Prepare the source plate for the chip Page 20 a Pipette cell samples into a 384 well sample source plate b Seal the plate with imaging film do not centrifuge 3 Dispense samples into the chip Page 20 a Perform chip focusing b Enter the chip ID into the software c Place the chip into the ICELL8 cx system d Place the sample source plate into the ICELL8 cx system e Dispense sa...

Страница 17: ...8 cx module c The live chamber temperature chamber relative humidity chuck setpoint and chuck temperature are displayed in the boxes below Figure 7 Example of DewPoint parameter display B Checking System Containers 1 Check the amount of water in the Water Bottle Make sure the Water Bottle is filled with Milli Q water to the 1 L mark on the bottle 2 Check the Humidifier Reservoir the one on NOT on ...

Страница 18: ...rmup each day prior to performing any experiments Failure to do so will result in poor dispensing This procedure takes 8 min 1 Click the Startup tab Figure 9 in the SMARTer ICELL8 cx Software Figure 9 Startup tab a On this tab click the Done button for Check water container Check humidifier water container Check waste container and Check bleach container Figure 10 Daily checklist ...

Страница 19: ... If bubbles are observed a Allow the Daily Warmup procedure to finish b Repeat the Daily Warmup procedure 4 Click System Prime on the Advanced tab a Click Wash Prime followed by Tip Clean if the Wash solution is refilled NOTE If large bubbles are still present after three Daily Warmup cycles we recommend priming the system tubing with isopropyl alcohol Refer to the GUI Startup tab Daily Maintenanc...

Страница 20: ...ng precautions when assembling sample and assay source plates Consider assembling source plates in a dead air box to reduce environmental dust Wipe down the lab bench every day and wear gloves and a clean lab coat Use plates tips and tubes from new or carefully covered containers Work quickly and cover plates tubes to minimize exposure to dust in the air 1 Pipette the cell sample for genotyping Co...

Страница 21: ...tretch the arms of the clip apart and angle the chip onto the Dispensing Platform with the beveled corner on the lower right and the edges of the chip pressed against the three alignment pins b Carefully release the clip that holds the chip in position Figure 14 Placing the chip on the Dispensing Platform C Placing the Sample Source Plate in the ICELL8 cx Unit 1 Place the sample source plate on th...

Страница 22: ... chamber has reached the RH and dew point setpoints D Dispensing the Sample Cells Fiducial and Controls into the Chip IMPORTANT Do not open the door of the cx Stage Module while the ICELL8 cx is dispensing If the door is open the chip can become contaminated Additionally when the door is open evaporation from the nanowells can occur resulting in changes to concentrations of the reagents in the chi...

Страница 23: ...io button will turn green If the chip is not secured the button will be red Figure 18 Vacuum status confirmation 4 If the chip is not secured you can take out the chip and restart the process by clicking the Dispense cells button again 5 The system will ask for confirmation on source plate and chip loading Confirm by clicking OK The dispense step lasts roughly 13 minutes Figure 19 Confirm plate an...

Страница 24: ... display for user reference Figure 21 Live image of chip scanning 8 8 The scanning process will take 1 2 minutes If the software is unable to read the chip ID manually enter the ID into the Chip ID field The dispense will proceed Figure 22 Input chip ID 9 When the dispense is complete the vacuum should turn off automatically and you can then release the chip Figure 23 Vacuum status after dispense ...

Страница 25: ...bio com Takara Bio USA Inc Page 25 of 35 E Imaging the Chip 1 When dispensing is complete press Scan chip to initiate scanning Figure 24 Initiate chip scanning 2 The vacuum will turn on automatically Remove the chip seal if necessary Figure 25 Remove the chip seal ...

Страница 26: ...barcode scanning steps are complete manually enter the chip ID in the Chip ID field if the software is unable to read the ID Figure 26 Enter the chip ID after autofocus and barcode scanning steps 4 Select the proper analysis setting and barcode gal files Press OK to confirm Figure 27 Select analysis setting and barcode gal files ...

Страница 27: ...me for saving data 6 Chip scanning will begin It takes roughly 13 minutes 7 When chip scanning is complete the system will ask if you want to save the result file Press Save to proceed 8 Name the results file Figure 29 Naming and saving results file 9 The vacuum should turn off automatically when scanning is complete You can remove the chip at this time 10 Close the SMARTer ICELL8 cx CellSelect So...

Страница 28: ...the blotter against the blotting paper on the chip The blotter should extend beyond the edges of the chip d Let the blotter rest on top of the blotting paper for exactly 2 sec without pressing down the weight of the blotter is sufficient for adequate blotting e Remove the blotter then gently remove the blotting paper and dispose of it in a biohazard container 2 Quickly seal the loaded chip with In...

Страница 29: ...ure 30 Cell RT dispense and scanning 2 Before the dispense step place the chip securely and flatly on the cool chuck and the loaded 384 well plate on the plate nest Refer to the plate map shown below Figure 31 Figure 31 RT PCR mix Source Plate map 3 Close the front door of ICELL8 cx unit 4 To initiate RT dispense click the Dispense RT PCR mix button Figure 32 Initiate RT dispense ...

Страница 30: ...filter files 6 The vacuum holding the chip will turn on automatically and the system will check the vacuum to confirm the chip sits flat If the chip is seated properly the vacuum radio button will turn green If the chip is not secured the button will be red 7 If the chip is not secured you can take out the chip and restart the process by pressing the Dispense cells button again ...

Страница 31: ...8 Manually remove the sealing film Click Done on the Remove seal window to confirm you have done so Figure 34 Confirm removal of sealing film 9 The system will perform autofocus and barcode reading steps The live image of chip scanning will display for user reference Figure 35 Live image of chip scanning ...

Страница 32: ...carefully remove it from the cooled chuck H Running your PCR on a Thermal Cycler If you are using the SMARTer ICELL8 cx Single Cell System Cat No 640188 use the thermal cycler Cat No 640002 included with the system See the thermal cycler s user manual for complete instructions on thermal cycling the filled chip If you are using the SMARTer ICELL8 cx Single Cell System JPN Cat No 640189 use a therm...

Страница 33: ...g Platform for any debris and wipe down with 70 isopropanol 2 Perform the Tip Clean procedure See Running the Tip Clean Procedure in Section VI D 3 Exit the SMARTer ICELL8 cx Software by clicking the Close button at the top right of the window 4 Turn off the ICELL8 cx unit by flipping the power switch on the rear of the box 5 Power down the computer that controls the ICELL8 cx 6 Remove all nanowel...

Страница 34: ... off a small section of the tube and reseat the tube Crimped fluidic tubing Inspect the tubing for crimps or bends Remove crimped bent sections if feasible or replace the harness Plugged tip Back flush the tips by clicking the Tip Clean button in the SMARTer ICELL8 cx Software If the tip is plugged with a soluble material aspirating and dispensing reagent capable of dissolving the material may cle...

Страница 35: ...A service technician or contact us directly at techUS takarabio com Contact Us Customer Service Ordering Technical Support tel 800 662 2566 toll free tel 800 662 2566 toll free fax 800 424 1350 toll free fax 800 424 1350 toll free web takarabio com web takarabio com e mail ordersUS takarabio com e mail techUS takarabio com Notice to Purchaser Our products are to be used for research purposes only ...

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