1.3.1 Hardware options
firefly instruments are available in the following configurations:
| Dispense heads | Shaker module | Thermal modules | |
| Genomics | 6 | Yes | Yes |
| Genomics | 3 | Yes | Yes |
| Non-genomics | 6 | No | No |
| Non-genomics | 3 | No | No |
The pipetting head options are the same for all instruments.
Illustrations in this manual show either the 6 head Genomics or the 3 head Non-Genomics
instrument.
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.
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 strips 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 (see section 7.5.7)
firefly loads clean tips automatically. To prepare the tips for loading (see section 6.4.1), 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 (see section 6.6.1).
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.
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.
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.
INFORMATION: 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 done off-line with an external thermocycler.
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.
1.3.8 Passive reagent cooling (all instruments)
To cool a single reagent, you could use a LDV reservoir and SPT Labtech's LDV reservoir cooling block. This is a passive cooling system, so you will not be able to control reagent temperature 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 Magnetic block
firefly is initially configured to work with Alpaqua and Permagen magnetic blocks for use with 96 well plates.
The Alpaqua Magnum FLX magnetic block is designed for fast magnetic bead separation. The magnetic block is compatible with standard 96 well PCR plates and deep well plates, with reaction volumes as large as 2 mL and elution in as little as 10 μL.
The Permagen 96-Well Ring PCR Magnet Low Elution Plate RLE500 is designed for use in automation applications where volumes as low as 5 μL are required. It has an angled frame around the top which helps to straighten PCR plates and locate them correctly on the sprung magnet.
If you need to use alternative magnetic blocks, check if they are available through Community Labware (see section 8). If they are not, and you need the specific type, contact reliance (see section 13) for advice.