Singer Instruments ROTOR HDA User Manual

Download

Page 1

USER GUIDE v1.2

Page 2

Before you start!

· All on-screen instructions MUST be followed. Straying from the

on-screen instructions could cause damage to the machine and the user.

· Ensure to keep transit hardware in a safe place. These will be

required should the ROTOR HDA require moving.

· During Pad Head removal, always ensure the push cylinder is cleared before moving away from the carriage. The push cylinder

runs from the carriage arm into The Stinger/Pad Head. If either are not pulled down far enough during removal, the cylinder will catch and damage the ROTOR HDA.

· Always ensure the power is OFF before changing heads. Leaving

the power on can cause a hardware crash.

Page 3

Anatomy and features

Removables

RePads

Mechanical overview

10.

Initialisation routine

12.

Software overview

14.

Program overview

16.

Program completion

17.

Software icon guide

22.

Technical specifications

24.

Pinning examples

25.

Post experimental procedure

Notes

26.

™

rotor HDa™

INTRODUCTION

The Singer ROTOR HDA

ultra-fast manipulation of high-density arrays of yeast, other fungi and bacteria. Reagent sets such as deletion mutant collections and the complete set of cloned yeast genes can be utilised for highthroughput screens; large-scale 2-hybrid, synthetic genetic array, phenotypic and chemical-genetic analysis. The ROTOR HDA uses plastic replica plating pads and supports liquid pinning to and from 96 and 384-well microtitre plates and agar pinning at densities of 96,

192, 384, 768, 1536 and 6144.

USER GUIDE

Follow these instructions alongside the on-screen instructions to get the most out of the ROTOR HDA, High Throughput Screening Robot.

This guide outlines basic operation of the ROTOR HDA as well useful

maintenance advice. Read through this guide and you’ll be ready for the exciting world of high throughput screening!

DISCLAIMER

At Singer Instruments, we are constantly seeking to improve our

products and adapt them to the requirements of modern research techniques and testing methods. This involves modification to the mechanical structure and optical design of our instruments.

Therefore, all descriptions and illustrations in this user guide,

including all specifications are subject to change without notice.

™

is a compact benchtop robot for easy,

singerinstruments.com

Page 4

anatomy & features

Dump Zone

Screen Handle Lid Lifters

Protective Screen

Fast Buttons

MCI Touch Screen

Carriage Arm

RS232 Shuttle Input

Emergency Stop

Hopper Loading Bay

Black BayDump Drawer

UV Decontamination Roll-over Cover

Turntable

Pad Head

Hopper

Control Panel

Page 5

anatomy & features

Beer Bottle Opener

Air Line Connection

Removable Transit Clamps

Removable Transit Bar

NOTE: Ensure to keep transit

hardware in a safe place should the ROTOR HDA require moving.

MCI Mounting Bracket

Power Switch

singerinstruments.com

Power In

Page 6

removaBles

BLACK BAY

· The Black Bay is a loading bay for source and target plates.

You will need to remove the Black Bay to load plates.

· If you own The Stinger single colony picker then you will

need to remove the Black Bay in order to change between the Pad Head and The Stinger.

PAD HOPPER

· The Pad Hopper is where the fresh RePads™ are loaded.

· Load a stack of RePads™ (pins facing down) into the Hopper

as shown, ensuring the chamfered corner is to the top right.

· The Pad Hopper is fully autoclavable to ensure RePads™

stay sterile. However, a pack of RePads™ protective card that means the RePads™ stay sterile without autoclaving the Pad Hopper.

includes a

DUMP DRAWER

· The Dump Drawer is where used RePads™ are deposited.

· When a program is finished, the Dump Drawer can be

removed to dispose of the used RePads™.

· The Dump Drawer is fully autoclavable.

Page 7

repaDs

™

96 SHORT

96 LONG

384 SHORT

COMPATIBLE MEDIA

· Solid agar

COMPATIBLE PLATES

· Liquid

· Solid agar

COMPATIBLE PLATES

· Solid agar

COMPATIBLE DENSITIES

· 96

· 192

· 384

· 1536

COMPATIBLE DENSITIES

· 96

· 192

· 384

· 1536

COMPATIBLE DENSITIES

· 384

· 768

· 1536

· 6144

384 LONG

1536 SHORT

6144 SHORT

COMPATIBLE PLATES

· Liquid

· Solid agar

COMPATIBLE PLATES

· Solid agar

COMPATIBLE PLATES

· Solid agar

COMPATIBLE DENSITIES

· 384

· 768

· 1536

· 6144

COMPATIBLE DENSITIES

· 1536

· 3072

· 6144

· 24576

COMPATIBLE DENSITIES

· 6144

· 12288

· 24576

singerinstruments.com

Page 8

mecHanical overview

1 2

Source Plates or Target Plates are loaded into the Black Bay. Source plates are the plates that already have the

desired strains on. Target plates are the plates that you want your desired strains to grow on.

· RePads™ are loaded (pins facing down) into the Pad

Hopper. RePads™ come in a variety of densities and are used to transfer strains from Source Plates to Target Plates.

· Target Plates or Source Plates are loaded into the Turntable.

· If loading four plates, the front two plates are loaded first.

The turntable will then rotate to allow you to load the second two.

· The Pad Hopper is loaded into the Hopper Loading Bay.

· The Pad Head lowers and picks up a RePad™.

· The Pad Head moves to the Source Plate, lowers and

collects a sample of cells.

Page 9

mecHanical overview

7 8

The Pad Head moves to a Tar get Pl ate and deposits the sample of cells.

9 10

· Used RePads™ are collected in the Dump Drawer ready to be

disposed of.

· These steps will be repeated until your chosen protocol is

finished.

· The Pad Head moves to the Dump Zone and drops the

used RePad™.

· Follow the simple step-by-step instructions for loading/

unloading of plates and consumables. See page 14.

singerinstruments.com

Page 10

initialisation routine

· Open the Roll-over Cover.

· Insert the Power Cables into the ROTOR HDA and

switch on the mains power.

· Turn on the ROTOR HDA Power Switch.

· Follow the simple on-screen instructions to work

through the ROTOR HDA initialisation procedure.

· Turn on the Compressor.

· Select OK on the Touch Screen or use the Fast Buttons.

Cancel

Page 11

initialisation routine

· Clear all objects from the ROTOR Stage, so nothing

will obstruct the initialisation routine.

· Select OK On-screen or use the Fast Buttons.

· The ROTOR HDA will perform a start-up routine.

singerinstruments.com

Page 12

software overview

This is the ROTOR Home Menu. From here you can choose to run pre-existing programs or create your own programs. In this example, we click Select And

Run Stored Programs.

· NOTE: Follow the same 4 steps to select for an existing

program:

1. Choose the type of Source Plates.

2. Choose the type of Target Plates.

3. Choose the type of Pin Pads.

4. Select required Program.

These four steps are demonstrated below.

· Select your Source Plates. They are the plates that

already have the desired strains on. The source could be colonies grown on solid agar, or cultures grown in liquid media. In solid agar, the ROTOR HDA currently supports colony density up to 24576. In liquid media, the ROTOR HDA supports multi-well plates at 96 and 384-density.

· In this example we select 96 Agar.

Select your Target Plates. They are the plates that you want your desired strains to grow on. The types of target plates available are only those compatible with your previous selection. Since we’ve selected our source plate as 96 Agar, we won’t be able to choose the target plate density beyond 1536.

· In this example we select 96 Agar.

· NOTE: The ROTOR software will not allow you to

make a mistake during plate and RePad™ selection.

Page 13

software overview

Select your RePad™. The types of RePads™ available are only those compatible with your previous selection.

· In this example we select Short Pin Pad 96-Density.

· NOTE: Short pin pads are used to pin colonies from

solid agar to solid agar. Long pin pads are used to pin liquid media or solid agar to solid agar or liquid media.

· The final step is to select the program that you want

to perform. The programs available are only those compatible with your previous selection. In this example, there are three available programs:

· Mate - pinning two haploid cells onto one plate.

· Replicate – pinning one source plate onto one target

plate.

· Replicate Many – pining one source plate onto multiple

target plates.

· In this example, we select Replicate Many.

· The right-hand panel will display your custom

programs. You also have the option to create new custom programs.

singerinstruments.com

Page 14

program overview

This is the Program Preparation Page. Here you can fine-tune the program using the options on the left. A detailed overview of these options can be found on pages 18-21.

· Select OK On-screen or use the Fast Buttons to

continue.

NOTE: Running a program is easy. Just follow the on-screen instructions!

Simple, step-by-step instructions will guide you through the program. Each plate position is colour coded. Red, blue, yellow and green go into the ROTOR Turntable, while black goes into the ROTOR Black Bay.

· Select OK On-screen or use the Fast Buttons when you

are done.

· When prompted, remove the Pad Hopper to load the

RePads™.

Page 15

program overview

Follow the instructions and load the appropriate

RePads™ into the Pad Hopper.

The ROTOR HDA will now count the number of Pin Pads loaded to ensure there are enough to complete the program.

· The ROTOR HDA will now start pinning the colonies for

replication.

· Live program information will be updated throughout

the operation.

· Quick button operation: Pause and Resume.

· On-screen operation: Resume, pause, stop / abort.

singerinstruments.com

Page 16

program completion

If you wish to generate more than 4 replicates of a target plate, you can repeat the program with one click.

· Make your selection.

· When pinning is completed, you will be instructed to

remove the Plates.

· Press OK when you are done.

· A Program Summary Page appears once the program

has completed.

Page 17

software icon guiDe

HOME SCREEN

· Home: This will return you to the ROTOR Home Screen.

· Unload: This allows you to remove the Hopper from the

Home Screen.

· Rotate: This rotates the Turntable.

· Reset: This resets the ROTOR HDA.

LOADING PLATES

· UV: This opens the UV Lamp Options.

· Info: This opens the Info and Settings

Screen. Here you can access the

advanced options and online support.

· Off: Press to turn off the ROTOR HDA.

· Load Plate

· Swap Plates

· Remove Plate

singerinstruments.com

Page 18

software icon guiDe

ADVANCE OPTIONS

ICON NAME SETTING DESCRIPTION TAB UNIT MIN MAX DEFAULT

Recycle

Revisit Source

Plate Protection

Off

Full

Until Repeat

During Pairs

On/Off

RePads™/pins are always dumped.

RePads™/pins are recycled for the duration of the program.

RePads™/pins are recycled for the duration of one cycle of the program.

RePads™/pins are recycled for the duration of each pinning pair.

Revisit ensures that the source is revisited for each pinning. If Revisit Source if off, the source plate will not be revisited unless a new position on the source plate is being pinned.

By protecting the source plates you can ensure that lids are only removed when it is vital to do so. This will increase the time it takes to run each program, but each source plate will be exposed for less time, and the print head will never move over a source plate without a lid on, unless it is pinning from it

General>Recycle N/A

General>Recycle

General>Plate Protection

N/A

Boolean

N/A

Off

N/A

Off

Repeat Pairs

Offset

On/Off

Off

Automatic

Random

A pinning pair represents pinning from a source plate to a target plate. You can adjust how many times each of these pairs are repeated. During pair repetition, Recycle and Revisit mode rules will be followed as normal.

No offset is used for source pinning.

An automatic offset is used for each source pinning.

A random offset is used for each source pinning.

General>Pairs

Source>Offset

Boolean

Off

Page 19

software icon guiDe

ICON NAME SETTING DESCRIPTION TAB UNIT MIN MAX DEFAULT

Offset

Source Pinning Pressure

Source Pinning Speed

Source Pinning Overshoot

Repeat Source Pinning

Manual

Fixed

Agar

Select a manual offset before each source pinning.

A pre-specified fixed offset is used for selected source pins.

The pressure that the Pad Head will use to push onto the agar.

The speed that the Pad Head will use to connect to the agar surface.

The amount of travel that will be applied after detecting agar contact. This is to enable operation of the pressure cylinde r.

The number of times each source pinning will repeat.

Source>Offset

Source>Pinning

Boolean

mm/s

Integer

Off

Speed

dependant

100

Speed

dependant

Off

Varies for each pad

Source Pinning Speed

Source Pinning Backoff

Repeat Source Pinning

Dry Mix Source

Dry Mix Clearance

Dry Mix Diameter

Liquid

On/Off

The speed applied to pinning to wet source plates.

The retraction distance applied to the Pad Head after sensing the bottom of the plate.

The number of times each source pinning will repeat.

Skipping around on the agar surface to select from a wider area of cells.

The distance the pins retract from the agar surface.

The diameter of the mix.

Source>Pinning

Source>Dry Mix

mm/s

Integer

Boolean

-0.5

Off

0.1

0.5

Off

0.5

Dry Mix Cycles

The number of cycles (comprising of 5 steps) that the dr y mix on source plates will be executed.

Source>Dry Mix

Integer

singerinstruments.com

Page 20

software icon guiDe

ICON NAME DESCRIPTION TAB UNIT MIN MAX DEFAULT

Wet Mix Source

SETTING

On/Off

Liquid mixing can be used to invigorate the cells in a liquid solution. Liquid

Source>Wet Mix

Boolean

Off

mixing uses either a circular or helical movement.

Source Mixing

The diameter of the mix applied to both the x and y axis.

Source>Wet Mix

Diameter

Source Mixing

The speed at which wet mixes are carried out.

Source>Wet Mix

mm/s

Speed

Source Mixing

The number of cycles the mix will include.

Source>Wet Mix

Integer

Cycles

Source Mixing

The distance that the Pad Head retracts on 3D mixes.

Source>Wet Mix

0.25

Trav el

Permanent offset

A permanent offset can be specified to reset the nominal centre for each

Source>Permanent Offset

Point

-3,-3

3,3

0,0

source pinning. This feature is useful when pinning from source plates that have been printed to a non-central location.

Tar get Pinning Pressure

Tar get Pinning Speed

Tar get Pinning Overshoot

Repeat Tar get Pinning

Tar get Pinning Speed

Tar get Pinning Backoff

Agar

Liquid

The pressure that the Pad Head will use to push onto the agar.

The speed that the Pad Head will use to connect to the agar surface.

The amount of travel that will be applied after connection to the agar surface has been made.

The number of times each target pinning will repeat.

The speed applied to pinning to wet source plates.

The retraction distance applied to the Pad Head after connection to the bottom of the plate.

Target>Pinning

Target >Pinning

mm/s

Integer

mm/s

Speed

dependant

-0.5

100

Speed

dependant

Varies for each pad

0.5

Page 21

software icon guiDe

ICON NAME DESCRIPTION TAB UNIT MIN MAX DEFAULT

Repeat Tar get Pinning

Dry Mix Tar get

Tar get Mixing Clearance

Tar get Mixing Diameter

Tar get Mixing Cycles

SETTING

Liquid

On/Off

Agar

The number of times each target pinning will repeat.

Agar mixing can be used to ensure that a good contact with the target media is established. Agar mixing prints multiple times at a specified radius around the target spot on the agar after the initial central print has been established.

The distance the Pad Head retracts from the agar surface at each stage of the mix.

The diameter of the mix. Using step-in reduces the diameter from the specified diameter uniformly with each cycle for a thorough mix.

The number of cycles (comprising of 5 steps) that the dr y mix on source plates will be executed.

Target >Pinning

Target >Dry Mix

Integer

Boolean

Integer

Off

0.1

Off

0.5

Wet Mix Tar get

Tar get Mixing Diameter

Tar get Mixing Speed

Tar get Mixing Cycles

Tar get Mixing Trav el

Pad Pickup Pressure

On/Off

Liquid

Liquid mixing can be used to ensure thorough depositing of cells in the liquid solution. Liquid mixing uses either a circular or helical movement.

The diameter of the mix applied to both the x and y axis. Using step-in reduces the diameter from the specified diameter uniformly with each cycle for a thorough mix.

The speed at which wet mixes are carried out.

The number of cycles the mix will include.

The distance that the Pad Head retracts on 3D mixes.

The pressure applied by the Pad Head when picking up pads.

Target >Wet Mix

Pads

Boolean

mm/s

Times

Off

0.25

100

Off

singerinstruments.com

Page 22

tecHnical specifications

DIMENSIONS

· Length: 1300mm (51”)

· Width: 650mm (26”)

· Height (from bench top): 725mm (29”)

NOTE: An additional 500mm (20”) is needed at one end for the bracket mounted MCI. This can fit at either end. The working height of the ROTOR turntable is 300mm (12 ”) from the benchtop.

NOTE: For servicing, the ROTOR HDA will require reasonable free space all round.

WEIGHT

· 110kg (242 lbs)

COLOURS

· Externally: White (with red Singer logo)

· Roller Cover: Grey

· Internal: White/grey

· Stations: Red/blue/yellow/green/black

NOCOST ACCESSORIES:

· Beer bottle cap remover or Corkscrew (specify).

POWER REQUIREMENTS

· 110-240V AC 50-60Hz Power: 500W

· Power connection at Right Hand End (from front) via IEC

Cable (supplied).

COMPRESSED AIR REQUIREMENT

(FOR COMPRESSOR, SEE PAGE 23)

· Dry, oil-free, compressed air/nitrogen at min 4 bar (60 psi)

max 10 bar (15 0 psi)

· Consumption: 3 l/min ( 0 .1 CFM)

· Air connects to LH end (from front) see Compressor Section

for connection details.

HEAD

· Movement: X: 800mm

Y: 30mm Z:90mm

· Clearance above table: 95mm

· Resolution: X: 1

Y & Z: 5

· Speed: X: up to 5,000mm/sec,

Y & Z: 25mm/sec (selectable)

· Control: X axis is closed-loop, linear motor with

linear encoder. Y & Z are open-loop stepper motor drives with optical data setting.

PAD HEAD

· Vacuum-operated and fully floating to comply with agar

surface.

· Programmable, variable pressure.

PAD DISPENSER

· Holds max 4 long-pin RePads™ and max 30 other types.

· Dispenser automatically counts RePads™ and flags up

shortage on GUI.

· Pad Dispenser rim fingers ensure accurate and repeatable

RePad™ positioning.

TURNTABLE

· Diameter: 360mm

· Angle of rotation: 180°

· Time: 2.5sec

· Repeatability better than 10

· Fitted with fully automatic plate positioners and latches.

· Bays are colour coded.

LID REMOVERS

· Triple, pneumatic, lift-and-turn lid removers each with

double hold-and-lift, vacuum-operated suction cups.

· Arms fitted with anti-rebound dampers.

FUNCTIONALITY

· Suspension transfer (wet/wet)

· Spotting (wet /dry)

· Colony replicating

· Array generation

· Mating

· Archive (dry/wet)

· Dry spotting (dry/dry)

DENSITIES OF MEDIA SUPPORTED

· 96, 384, 1536 and 6144 RePads™ (solid agar)

· Long-pin 96 and 384 RePads™

solid/liquid)

PL AT E

(liquid/liquid-liquid/solid -

· 96 and 384-well footprint, standard depth

· Singer PlusPlates™ (Rectangular, single extended cavity, 96-

well footprint)

MACHINE CONTROL INTERFACE

· 15” touchscreen 1224 x 788 resolution

· Intel Atom Processor

· 1GB R AM

· 1.8GHz

· 10GB hard drive

· Windows XP embedded standard

Page 23

tecHnical specifications

GRAPHICAL INTERFACE

· All functions controlled by simple pictograms.

Includes replication, array generation and mating

SOFTWARE

· Commands include automatic and manual offset pinning

to ensure even repeat cell pick up from colonies and automatic stirring mode for re-suspention in microtitre wells. Remote access and diagnostics and other protocols are under continuous development.

INTERFACES

· 1x Ethernet

· 2x USB

· 1x 25232

· 1x KB/MS/LAN2

LIGHTING/DISINFECTION

· White

· UVc

COMPRESSOR

· Compressor type may vary, please consult your Singer

Technician.

· Our standard compressor is very quiet and performs

optimally standing on the floor. It has a reservoir inside it and will run only intermittently. The pipe connecting the compressor to the ROTOR HDA is 6mm dia (1/4”). The compressor may be sited away from the ROTOR HDA

(please let us know about this so that we can supply a long enough pipe).

· 120V or 230V versions of our standard compressor are

available.

· Power: 500W.

AIR/GAS CONNECTION

Where we do not supply a compressor, the ROTOR HDA is supplied with a male quick-change coupler. We will supply, in advance, the female mating part to this, so that you can arrange connection before installation.

PERFORMANCE

The ROTOR HDA is manually loaded and unloaded, but very special attention has been paid to speed of replication. The turntable, which conveys plates in and out of the sealed operating zone of the ROTOR HDA, may be unloaded and loaded whilst replication is in progress. This makes the process very continuous.

Performance tests carried out for Singer by a major yeast laboratory claim replication rates in excess of 100 PlusPlates™ per hour.

At the supported densities, this equates to:

· 96: 9,600 colonies

· 384: 38,400 colonies

· 768: 76,800 colonies

· 1536: 153,600 colonies

· 6144: 614 ,40 0 colonie s

CONSUMABLES

· Singer PlusPlates

plate with specially extended working area for RePad compatibility and meniscus allowance.

™

: Standard footprint, single well

™

· RePads™: 96 Long

384 Long 96 Short 384 Short

1536 Short

6144 Short

All consumables are made of plastic and are gamma irradiated and double packed.

Pack sizes:

· PlusPlates™: 10 per sleeve/ 200 per box

· Long 96: 10 per sleeve/ 200 per box

· Long 384: 10 per sleeve/ 200 per box

· Short 96: 20 per sleeve/ 1000 per box

· Short 384: 20 per sleeve/ 1000 per box

· Short 1536: 20 per sleeve/1000 per box

· Short 6411: 20 per sleeve/1000 per box

Running Costs

· Running costs of the ROTOR HDA are much lower than

those of conventional robots, particularly when speed and density are taken into account.

· EXAMPLE: At a rate of 100 plate replications per hour

at a density of 1536, 153600 colonies can be printed. 100 PlusPlates™ and 100 RePads™ cost approximately US $240* so cost per colony is only $0.0015. There are no solvent or detergent costs, or costs associated with a dedicated technician.

SAFETY AND COMPLIANCE

· Safety Interlocks on vision panel and rear control panel.

· UV lamp operation is under software control and is

interlocked with main, roller cover closure.

· CE compliance by technical file.

*Price correct at time of printing. Exchange rates fluctuate monthly.

singerinstruments.com

Page 24

pinning examples

1:4 A R R AY

· 4x 96-density plates are combined onto

1x 384-density plate.

1:4 SINGLE SOURCE

· Each colony from a 1x 96-density plate

replicated in quadruplicate to a 1x 384density plate. These protocols can be applied at all pinning densities.

1:4 A r r ay

1:4 Single source

Page 25

post-experimental proceDure

1 2

Select a Delicious Beer. Pick a strong one - it’s been a

hard day!

· Insert the Beer Bottle into the Bottle Opener.

· Locate the Bottle Opener on your ROTOR HDA.

· Lever the Beer Bottle to remove the Bottle Cap

· Place the Bottle Cap in the Bin. Nobody likes a litter bug!

· Success! Time to enjoy your Delicious Beer - you’ve

earned it!

· Repeat steps 1-6 until suitably relaxed.

singerinstruments.com

Page 26

notes

Page 27

notes

singerinstruments.com

Page 28

notes

Page 29

notes

singerinstruments.com

Page 30

Ro adw ater, Watchet, Somerset. TA23 0RE UK

+44 (0)1984 640226 (tel) +44 (0)1984 641166 (fax)

contactsingerinstruments.com singerinstruments.com

scan to vi sit we Bsite for mo re

Helpf ul tip s anD t uto ria ls!