iota Sciences isoCell Manuale utente
V1.0r1
isoCell user manual
For software versions 1.0.0 - 1.0.8
Software v1.0.7 release notes
From software version 1.0.7 onwards, the total volume of fluid in the
chambers has been reduced from 800nl to 600nl. This will provide greater
chamber stability. Chambers are still plated with 200nl, but Fill will now only
add 400nl to the chambers (rather than 600nl). Other programs have been
modified to work with the lower volume chambers, but are otherwise
unchanged.
›
isoCell Manual
isoCell Manual
Version
Version v1.1
v1.1
Introduction
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.
Safety 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.
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:
Use only with the supplied
Mean Well GST90A24-P1M
power supply and power
cord.
Connect power supply to a suitable AC mains supply with voltage in range
100 – 240V; 50 – 60Hz; minimum power 1A.
The AC mains socket shall act as the disconnect device and be readily
accessible.
Avoid subjecting the isoCell to impact and vibration, both during use and during
transportation.
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.
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 designed to work between 16 – 35°C (60 – 95°F) and 40 – 60%
humidity. Operating outside these limits may degrade performance and
damage the isoCell.
All service shall be performed by iotaSciences authorised personnel only.
Getting started
Familiarisation
Users should familiarise themselves with the key components of the isoCell (Figure 1)
prior to use.
Figure 1: Key components of the isoCell.
N.B. the selection pillar is by default undeployed and therefore may not be visible.
The isoCell is supplied with the following accessories, consumables and spares:
Documents:
isoCell quickstart user guide
isoCell training guide
Single cell suspension preparation guidelines
Mains power cable, with appropriate mains plug for country of use
Transformer (24V Mean Well GST90A24-P1M power supply)
USB cable, type A – type B
FC40STAR bottle assembly, including 2-port cap and 0.20µm air filter
Touchscreen stylus
Reference stage
50-page single-cell microscope notebook
2x spare jetting assembly
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 10-page single-cell microscope notebook
1x 40ml bottle of FC40STAR
Installation
The isoCell should be located on a flat, stable surface inside a laminar flow hood.
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.
Using the isoCell
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).
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.
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:
The isoCell will provide a visual prompt of the reagents required by any given
part of the workflow.
A seventh reagent, FC40STAR, is stored in the FC40STAR bottle at the rear of the
isoCell (Figure 1).
Before beginning any program, ensure all microcentrifuge tubes are open
with their lids tucked under the rails, as shown in Figure 2. This is essential to
prevent obstruction of the needle holder.
Two further (empty) microcentrifuge tubes serve as repositories for aqueous and
FC40STAR waste. The isoCell will provide a visual prompt when waste microcentrifuge
tubes are required. Users should ensure that waste tubes are empty.
For instructions on the safe disposal of FC40STAR, refer to
Disposal of FC40STAR
.
Cells are harvested by the isoCell from the 60mm dish into the two 8-tube PCR strips
(or flat bottom 8-well strips) located at the front of the bed (Figure 2). The isoCell will
provide a visual prompt when the PCR or flat bottom strips are required. The strips
should be prepared with 120µl cell culture medium in each tube/well.
Do not put caps on the PCR strips.
Running programs
The isoCell workflow of isolating, culturing and harvesting cells is executed by
running a series of programs on the isoCell. For instance, the
GRID
program will
create a GRID on the 60mm dish, the
plate
program will dispense cells into a GRID,
etc. All programs are executed from the isoCell’s interactive touchscreen. A stylus is
supplied for higher touch accuracy when required.
At the beginning of each program, a message on the touchscreen will prompt the
user to place the correct materials and reagents onto the
isoCell bed
. When
providing reagents in microcentrifuge tubes, the user should fill the tube to a
minimum volume of 1ml. When a waste tube is required, the user should ensure that
the tube is empty.
Many programs require the user to select a username and dish colour. The
isoCell supports up to thirty-six users. Under each username the isoCell can
simultaneously track up to nine individual dishes via the dishes’ coloured
locating rings. Stored user data can be managed via the
data
program.
Starting-up
Upon being powered-on, the isoCell will execute a startup routine.
Users should not skip the startup routine.
Restarting
To restart the isoCell, first press the on-screen button (located on the left side of
the touchscreen). Then select the
restart
option, which will initiate a restart routine.
The restart routine will put the isoCell into a fresh state. It is recommended
that when switching users, the new user restarts the isoCell in this way.
Powering-off
To power-off the isoCell, first press the on-screen button (located on the left side
of the touchscreen). Then select the
shutdown
option, which will initiate a shutdown
routine. At the end of the shutdown routine, the isoCell will indicate that power can
be safely switched off. At this point the user should power-off the isoCell by pressing
the power button (located on the top of the isoCell).
Users should not skip the shutdown routine.
Consumables ordering
To order consumables for the isoCell (CloneG kits and FC40STAR), please visit the
consumables store on the iotaSciences customer portal to obtain an instant quote.
Workflow
The isoCell offers a convenient and user-friendly workflow to achieve consistent and
reliable outcomes from single-cell cloning experiments. It enables users to:
1. Isolate single cells
2. Culture and outgrow single cells into populations/colonies
3. Harvest populations/colonies derived from a single cell
The isoCell supports up to thirty-six users. Under each username the isoCell can
simultaneously track up to nine individual dishes via the dishes’ coloured locating
rings.
The
isoCell quickstart user guide
provides a concise overview of the isoCell
workflow. The
isoCell training guide
provides a more detailed introduction to
the isoCell workflow for new users. Both documents are supplied with the
isoCell, and available for download from the iotaSciences customer portal.
Isolate
Cells are isolated in three stages:
1. The
GRID
program is used to form a 256 chamber GRID on a 60mm tissue culture
dish, with each chamber measuring 1.9mm by 1.9mm.
2. The
plate
program is used to dispense 200nl cell-suspension fluid to each of the
GRID’s 256 chambers.
3. The
input data
program is used to store, in the isoCell’s memory, the GRID-
coordinates of chambers containing single cells.
GRID
To form a GRID, the user must first prepare the tissue culture dish with a coating of
cell culture media/supplement formulation, overlaid with FC40STAR.
Detailed dish-preparation protocols for specific cell culture
media/supplement formulations are available on the iotaSciences customer
portal.
For successful GRID formation, ensure that the relevant dish preparation protocol is
followed correctly. Once prepared, place the dish onto the isoCell bed and run the
GRID
program.
Plate
To plate cells, the user must first prepare a 1.5ml microcentrifuge tube containing a
sample of cell-suspension fluid with a dilution ratio of 5000 cells per millilitre.
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