IOTA isoCell User manual

v2.1r4
isoCell user manual
For software versions 2.1.0 and 2.1.1

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Products bearing the symbol of a strikethrough refuse bin must not be disposed of
along with unsorted refuse.
For details on how to dispose of your isoCell at the end of its useful life, see section
8.1.
NOTE: This equipment has been tested and found to comply with the limits for a Class B digital
device, pursuant to part 15 of the FCC Rules. These limits are designed to provide reasonable
protection against harmful interference in a residential installation. This equipment generates,
uses and can radiate radio frequency energy and, if not installed and used in accordance with
the instructions, may cause harmful interference to radio communications. However, there is no
guarantee that interference will not occur in a particular installation. If this equipment does
cause harmful interference to radio or television reception, which can be determined by turning
the equipment off and on, the user is encouraged to try to correct the interference by one or
more of the following measures:
—Reorient or relocate the receiving antenna.
—Increase the separation between the equipment and receiver.
—Connect the equipment into an outlet on a circuit different from that to which the
receiver is connected.
—Consult iotaSciences for further help.
FCC CAUTION: Any changes or modifications to the isoCell other than those shown in the
manual, demonstrated during training, or directed by iotaSciences authorised personnel may
void compliance and the user’s authority to operate the isoCell.

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Contents
1 Introduction.........................................................................................................................................................................................................................................5
2 Safety instructions.......................................................................................................................................................................................................................5
3 Getting started.................................................................................................................................................................................................................................6
3.1 Familiarisation......................................................................................................................................................................................................................6
3.2 Installation................................................................................................................................................................................................................................8
3.3 Using the isoCell...............................................................................................................................................................................................................9
3.3.1 Powering-on............................................................................................................................................................................................................9
3.3.2 The isoCell bed –placement of reagents and materials .................................................................................9
3.3.3 Running programs..........................................................................................................................................................................................10
3.3.4 Starting-up..............................................................................................................................................................................................................10
3.3.5 Restarting................................................................................................................................................................................................................... 11
3.3.6 Powering-off..........................................................................................................................................................................................................11
3.4 Consumables ordering.............................................................................................................................................................................................11
3.5 Using the isoCell as part of the iotaSciences Cloning Platform.......................................................................... 11
4 Workflow...............................................................................................................................................................................................................................................12
4.1 Isolate ..........................................................................................................................................................................................................................................12
4.1.1 GRID.................................................................................................................................................................................................................................12
4.1.2 Plate.................................................................................................................................................................................................................................13
4.1.3 Input data .................................................................................................................................................................................................................13
4.2 Culture ........................................................................................................................................................................................................................................14
4.2.1 Fill........................................................................................................................................................................................................................................14
4.2.2 Feed.................................................................................................................................................................................................................................14
4.3 Harvest .......................................................................................................................................................................................................................................15
4.3.1 Detach...........................................................................................................................................................................................................................15
4.3.2 Extract...........................................................................................................................................................................................................................15
4.3.3 Redetach....................................................................................................................................................................................................................16
4.4 Chamber states................................................................................................................................................................................................................16
5 Advanced programs.................................................................................................................................................................................................................17

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5.1 Data................................................................................................................................................................................................................................................ 17
5.2 Replace dispensing......................................................................................................................................................................................................18
5.3 FC40STAR® level...................................................................................................................................................................................................................18
5.4 Prep transport...................................................................................................................................................................................................................18
5.5 Reset jetting.........................................................................................................................................................................................................................18
5.6 Reset dispensing ............................................................................................................................................................................................................19
5.7 Recover......................................................................................................................................................................................................................................19
5.8 Calibrate....................................................................................................................................................................................................................................19
5.9 Move.............................................................................................................................................................................................................................................19
5.10 Options..................................................................................................................................................................................................................................... 20
5.11 Set time.....................................................................................................................................................................................................................................21
5.12 Data sync.................................................................................................................................................................................................................................21
5.12.1 Setting up data syncing.........................................................................................................................................................................22
5.13 About ..........................................................................................................................................................................................................................................23
6 Maintenance.....................................................................................................................................................................................................................................23
6.1 Cleaning ...................................................................................................................................................................................................................................23
6.2 Replacing the FC40STAR® bottle....................................................................................................................................................................23
6.3 Replacing the dispensing assembly ..................................................................................................................................................... 24
7 Troubleshooting.......................................................................................................................................................................................................................... 24
8 Disposal and recycling .........................................................................................................................................................................................................25
8.1 Disposal of isoCell.......................................................................................................................................................................................................25
8.1.1 EU Waste Electrical and Electronic Equipment (WEEE) Directive.....................................................25
8.1.2 Users outside the European Union..........................................................................................................................................25
8.2 Disposal of FC40STAR® ...............................................................................................................................................................................................25
8.3 Disposal of reagents and consumables............................................................................................................................................25
9 Technical support.......................................................................................................................................................................................................................26
10 Additional information.................................................................................................................................................................................................26
10.1 Limited warranty...........................................................................................................................................................................................................26
10.2 Limited liability................................................................................................................................................................................................................26

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10.3 Medical use restrictions.......................................................................................................................................................................................27
10.4 Registered office...........................................................................................................................................................................................................27
10.5 Legal information.........................................................................................................................................................................................................27

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1Introduction
The isoCell greatly simplifies and automates the cloning of single cells, by allowing users to:
•Create GRIDs on 60mm circular dishes.
•Dispense single cells into GRID chambers (following the Poisson distribution).
•Add cell-culture medium to GRID chambers, exchanging for waste products.
•Harvest colonies.
2Safety instructions
Use the isoCell with care, and take the following precautions:
•Only use the isoCell once you have read and understood this manual. Ensure this manual is
accessible for all users, and that each potential user reads and understands it.
•If the isoCell is used in a manner not specified by the manufacturer, then the protection
provided by the isoCell may be impaired.
•Take all suitable precautions recommended for the safe handling of all cells and other
reagents used. For the case of FC40STAR®, users should familiarise themselves with the FC40
material safety data sheet, which is available on the iotaSciences customer portal.
•Powering the equipment:
oUse only with the supplied XP-POWER VEC50US24 (24VDC, 50W) power supply
and power cord.
oConnect power supply to a suitable AC mains supply with voltage in range 90 –
260V; 50 - 60Hz; minimum current 1A at 115V, 0.7A at 230V.
oThe AC mains socket shall act as the disconnect device and be readily accessible.
•Avoid subjecting the isoCell to impact and vibration during use.
•Keep the original packaging the isoCell came in, and use this when transporting the isoCell.
•Do not connect cables in ways other than those mentioned in this manual. Different
connections may cause damage and fire.
•Do not alter the isoCell in any manner, other than those shown in the manual,
demonstrated during training, or directed by iotaSciences.
•Do not obstruct the movement of the isoCell’s needle holder, nor of the syringe plunger
mounts.
•Only use the isoCell when it is set on top of a safe, flat and stable horizontal surface.
•If water or other liquid has been spilled near the isoCell, safety may be compromised.
Ensure that no spilled liquid has been in contact with or entered the isoCell.
•If water or other liquid enters the isoCell, stop using it immediately and unplug the power
supply. Continued use may cause a fire. Contact iotaSciences for actions or precautions.

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•If any foreign object enters the isoCell, safety may be compromised. Contact iotaSciences
for actions or precautions.
•Unplug the isoCell immediately if it smokes, or produces a strange odour or unusual noise.
Continued use may cause a fire. Contact iotaSciences for actions or precautions.
•Do not autoclave the isoCell.
•The isoCell is for indoor use only.
•The isoCell is designed to work in the following operating environment (operating outside
these limits may degrade performance and damage the isoCell):
oAmbient temperature: 16 –35°C (60 –95°F)
oHumidity: 40 –60%
oAltitude: less than 2000m
oPollution degree 2
•All service shall be performed by iotaSciences authorised personnel only.
•External influences such as electrostatic discharge may cause this equipment to
malfunction. In this case, please disconnect the power and reconnect.
3Getting started
3.1 Familiarisation
Users should familiarise themselves with the key components of the isoCell prior to use (Figure 1).

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Figure 1: Key components of the isoCell. (N.B. the selection pillar is by default undeployed and therefore may not be
visible..)

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The isoCell is supplied with the following accessories, consumables and spares:
Documents (also available for download from the iotaSciences customer portal.):
•Quickstart user guide
•Training guide
•Single cell suspension preparation guidelines
Mains power cable, with appropriate mains plug for country of use
Transformer (24V XP-POWER VEC50US24 power supply)
USB cable, type A –type B
Touchscreen stylus
Reference stage (not included if isoCell has been purchased as part of the
iotaSciences Cloning Platform)
50-page single-cell microscope notebook (not included if isoCell has been
purchased as part of the iotaSciences Cloning Platform)
2x 40ml bottles of FC40STAR®
CloneG consumables kit, comprising:
•18x 60mm dishes (either tissue-culture treated or suspension) with
attached locating rings
•36x eight-tube PCR strips, or 36x flat bottom 8-well strips
•50x 1.5ml microcentrifuge tubes
•1x pipetting aid
•2x dispensing assembly
Training kit, comprising:
•6x tissue-culture treated 60mm dishes with attached locating rings
•12x eight-tube PCR strips
•25x 1.5ml microcentrifuge tubes
•1x pipetting aid
•1x 40ml bottle of FC40STAR®
3.2 Installation
The isoCell should be located on a flat, stable surface inside a laminar flow hood.

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Upon first being powered-on, the isoCell will initiate a preliminary setup procedure. This consists
of a sequence of on-screen instructions for the user to follow, and requires approximately 30
minutes to complete. Upon completion, the isoCell will be ready to use.
3.3 Using the isoCell
3.3.1 Powering-on
To power-on the isoCell, ensure that it is connected to a suitable mains power socket by the
supplied mains power cable and 24V transformer. Press the power button (located on the top of
the isoCell).
3.3.2 The isoCell bed –placement of reagents and materials
The isoCell bed (Figure 2) is where the workflow of isolating, culturing and harvesting cells is
executed.
Figure 2: The isoCell bed, showing correct layout of 60mm dish, microcentrifuge tubes and 2x 8-tube PCR strips
(optionally replaced with 2x flat bottom 8-well strips).
The workflow is executed on a 60mm tissue culture dish (supplied in the CloneG consumables kit).
The dish must be placed in the circular station at the centre of the bed. It must be seated flat, and
oriented such that the triangular protrusion of the locating ring fits into the station’s triangular
notch (Figure 2).
The isoCell bed is equipped with an internal heater, which can be turned on or off in the
options program (Section 5.10). N.B. The heater is not intended to hold the dish at
precisely 37°C. Rather its purpose is to slow cooling when the dish is removed from an
incubator.
When required by the workflow, user-supplied reagents must be placed in 1.5ml microcentrifuge
tubes (supplied in the CloneG consumables kit) at six defined locations on the bed (Figure 2). A
minimum of 1ml of each required reagent should be provided. Note that: