1.3.1 Hardware options
firefly instruments s variants are available which use either ATL (Auto Tip Load) or EZL (EZ-Load) pipette tip technologies.
ATL instruments can be supplied with a firefly+ module for additional plate handling capabilities, and optional thermocycler capability.
| Dispense heads | Shaker module | Thermal modules | |
| Genomics | 6 | Yes | Yes |
| Genomics | 3 | Yes | Yes |
| Non-genomics | 6 | No | No |
| Non-genomics | 3 | No | No |
Illustrations in this manual show a variety of instrument types and configurations.
1.3.2 Dispensing
firefly syringes can be synchronized for high throughput dispensing or utilized independently for reagent flexibility. The positive displacement syringes dispense volumes from 200nL to 3690µL. Each 4mL syringe can be used to multi-dispense smaller volumes for faster dispensing to a 96 or 384 well plate. The non-contact dispensing mechanism enables syringes to be used for multiple
operations per reagent.
The syringe dispensing heads can dispense to plates located in deck positions 6 and 7, on both the upper and lower decks.
The syringes are filled from 6 reservoirs, located in a reservoir tray in the basement.
There are multiple reservoir and reservoir tray types available, to meet different dispensing needs.
1.3.3 Pipetting
firefly can pipette volumes from 0.5μL to 125μL.
The pipetting head has tip options to operate in several configurations: 384 and 96 tips for full plates, or columns of 16 (384 format) and 8 (96 format) strip tips, for working with partial plates or working across plates in processes such as serial dilutions.
The pipetting head can access all deck positions when using arrays of tips and columns of tips. There are spatial limitations associated with the movement of the pipetting head when relatively tall labware surrounds the pipetting location, but firefly software will warn you of these during the protocol design process.
There are two types of pipetting head.
EZL head
The EZL head uses a stainless steel cassette - visible above the tips in the illustration below - to hold pipette tips. Tips can be loaded as either a complete set of 96 or 384, or tip strips can be used for partial plate pipetting.
To prepare EZ-load tips for loading, you put them into a tip cassette which you place on a tip stand on the lower deck, and firefly software controls loading the tips to the pipette head. Tip removal is also automated: you simply need to unload the tip stands of used tips and then remove the tip cassette from the stand.
EZ-load firefly is supplied with 5 tip stands and 5 tip cassettes as standard. Additional tip stands, tip cassettes and strip tip adaptors are available from SPT Labtech.
ATL head
The ATL head tips are directly loaded from the tip box, with the quantity specified in the firefly protocol. You can load anything from a single column of tips to the complete box of 96 or 384 tips.
firefly loads clean tips automatically. To load ATL tips, you optionally place the tip box on a tip box adapter and firefly software controls loading the tips to the pipette head. Tip unloading is also software controlled; you simply load an empty tip box into which the used tips are unloaded, ready for removal and disposal. ATL firefly is supplied with 1 tip box adapter.
1.3.4 Decks and grippers
firefly has two movable decks, each with space for 8 plates.
The top deck can be used for items with a maximum height of 49.7mm. The bottom deck has a maximum height limit of 93mm so deep items such as tip sets must be loaded on the bottom deck.
The ‘Auto Fill Deck’ function (highlighted) in firefly software will determine a correct loading location for all the items used in a protocol.
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The pipetting head incorporates grippers to move labware on and off the plate thermal module and shaker module. It can move labware between decks and also on and off accessories such as magnetic blocks. Grippers can also be used to stack items e.g. a plate and a thermal block, and then move them together as shown below. On a Genomics instrument, grippers move labware on and off the plate thermal module and shaker module.
The gripper can move labware when tips are loaded onto the pipetting head as it reaches down below the tip level, or tips can be unloaded before working with the gripper.
1.3.5 Plate heating and cooling (Genomics option)
The plate thermal module provides plate heating and cooling. Temperatures are selectable from -20°C to 99°C in steps of 0.1°C. The nominal regulation accuracy is better than ±0.1°C with a uniformity of temperature distribution better than ±0.7°C at 4°C across the cooling surface.
Plate thermal module ramping times are rapid, with a cooling/heating speed 12°C / min above RT, but the module is designed for incubations only. PCR should be carried out off-line using an external thermocycler or the firefly+ ODTC.
1.3.5.i Thermal block
The plate thermal module is used with a thermal block, an adapter which is placed on top of the plate thermal module beneath the plate. Remove it from the plate thermal module with the plate and it will continue to provide passive heating or cooling: it will slow down the rate of temperature change of the plate contents to approximately 1°C per minute.
There are specific thermal blocks for 96 and 384 well plates. A block for 96 well PCR plates is supplied as standard. You can purchase additional blocks as you can concurrently use multiple blocks on an instrument:
- 3276-01065 THERMAL ADAPTER FOR PCR PLATE (96 WELL)
- 3276-01066 THERMAL ADAPTER FOR PCR PLATE (384 WELL)
1.3.6 Plate shaking (Genomics option)
The shaker module mixes at speeds between 200 - 3000 rpm.
The shaker can be used with all firefly plate types, including deep well plates although there is a height restriction meaning that decks could not be moved with some deep blocks on the shaker.
1.3.7 Reagent cooling and heating (Genomics option)
The dispense head reservoirs can be cooled by using the optional reservoir thermal module.
This has the same heating and cooling properties as the plate thermal module. Temperatures are selectable from -20°C to 99°C in steps of 0.1°C. All reservoirs are cooled or heated to the same temperature. If using the reservoir thermal module, you replace the standard plastic reservoir tray with an aluminum reservoir tray.
If you wish to use high volume reservoirs, you will need to use the specific reservoir tray (shown
below) and the inserts for standard or low volume reservoirs. Lids are available for high volume
reservoirs, to reduce evaporation.
1.3.8 Passive reagent cooling (standard reservoir tray)
To cool a single reagent, you could use a LDV reservoir and an LDV reservoir cooling block. This is a passive cooling system, so you will not be able to maintain reagent temperature or control it precisely.
- 1. Put the LDV reservoir cooling block in a freezer for 2 hours.
- 2. Place the chilled block in the reservoir tray beneath the LDV reservoir.
NOTE: As the chilled block is not an asset in firefly software, you will need to use user interactions or your method documents to include instructions to use it.
1.3.9 Passive reagent cooling (HV reservoir tray)
To cool a single reagent, use a standard reservoir and insert or a LDV reservoir and insert. The
aluminum inserts are shown below.
This is a passive cooling system, so you will not be able to maintain reagent temperature or
control it precisely.
- Put the insert in a freezer for 2 hours.
- Place the insert in the HV reservoir tray beneath the reservoir.
You could also use a pre-chilled insert in addition to the thermal module, e.g., if one material
needs to be at a lower temperature than the rest.
1.3.10 Labware barcodes
firefly software enables you to capture barcodes from your plates and reagents, and from firefly reservoirs, syringes and tips, using the handheld barcode reader. You can optionally include or
omit the use of barcodes in the protocols you design on an item by item basis e.g., you could just
record sample barcodes.
1.3.11 Magnetic block
You can use a variety of Alpaqua and Permagen magnetic blocks with firefly, for use with 96 and
384 well plates.
Before you can use a magnetic block, you must download the definition from Community
Labware. If you need a specific type which is not available, contact reliance for advice.
You will also need to download the stack definition for the magnetic block with the plate which
you want to use, otherwise it will not be available when you are designing your protocols
1.3.12 firefly+
firefly+ extends firefly's capabilities with a choice of integrated thermocycler modules and additional storage capability for plates, lids and tips. It is only available for use with the firefly ATL head variant.
It includes:
- An optional Inheco On Deck Thermal Cycler in 96- or 384-well format (if you don't require this, a drip tray is installed in the base of firefly+)
- 15 x medium shelves (max. labware height 67mm - ideal for additional tip boxes)
- 9 x small shelves (max. labware height 27mm - ideal for additional plates and lids)
The shelves are arranged to hold SLAS/ANSI (SBS) labware within the height limit of the particular shelf, as shown above.
Do not use firefly+ for storing filled trough plates as the movement of the firefly+ grippers is likely to spill the contents.
1.3.13 On Deck Thermal Cycler (ODTC)
The Inheco On Deck Thermal Cycler XL is a firefly+ option which enables you to carry out incubation steps in PCR, NGS and gene analysis protocols without requiring a separate instrument.
You cannot dispense or pipette directly to a plate while it is in the thermocycler, as it is situated in firefly+ and not accessible to the dispensing and pipetting heads.
Details of the ODTC and instructions for use are in the Inheco On Deck Thermocycler manual.
You can only use skirted PCR plates with the On Deck Thermal Cycler and the firefly+ grippers. While the ODTC is compatible with semi-skirted plates, firefly's grippers cannot pick these plates up to transfer them.
The maximum height of plates that can be placed on the ODTC is 22mm.
Next Article - Installation - 2.1 Pre-installation requirements