Evaluation and Implementation of

Machine-assisted Slide Processing

Charlotte GregorySouth-East Scotland Cytogenetics Service

Western General

Edinburgh

Time Consuming

Labour Intensive

Inconsistent

Messy

Slide Processing

Genetix SL-120

Metaphase Finder/Analysis

Transgenomics Hanabi

Automatic Harvester

Aims of the Project

• Integrate new equipment

(Set up/Programming/SOPS/Training)

• Compare quality and consistency to manual method

• Time, Space, financial implications

Thermotron (Fisher)

Environmental Chamber

• Controlled Environment

• Gives Consistency

• Facilitates Spreading

• Optimum values?

TemperatureOC

RelativeHumidity %

%Metaphase ≥ QA5

B F

AverageCrossovers

23.8 Control in Hood

45 23 27 8 7

20 50 50 50 9 6

25 50 49 56 5 6

30 30 50 33 9 8

Variables

•20, 25, 30oC

•20, 30, 40, 50 ,60% RH

for each temperature as above

Varistain Gemini (Shandon)

Automatic Stainer

• Robotic arm/troughs

• Multiple Samples

• Accurate Timing

• Heat/Running Water

Postnatal

Pre-treatment

Hot Plate/Peroxide

Time 1hr/30mins

TrypsinConcentration

1/10

Time 50s

PBS wash 5s + agitation

Leishman’sConcentration

1/4

Time 2.5m

Gurr’sWash

5 secs +agitation

•Postnatal

•Prenatal

•Tissue

•Haematology

•Tumour

Adapted from Manual Protocol

video

Picture of reagent screen

1m 1m 1m5m

Load Trypsin PBS Stain Gurr’s Store

1 rack every 8 minutes (32 minutes for 4)

Previous Version

Updated Version

Load Store Trypsin PBS Stain Gurr’s Store

1m 1m 1m

5m

5m

5m

5m

4 racks at same time (11 minutes for 4)

Picture of sample screen

Clearvue (Shandon)

Automatic Coverslipper

• Daily maintenance

• Preloaded Coverslips

• Humid atmosphere

• No pressure

video

Methods % Cells ≥ QA5

Spreading Banding

Thermo/Man 67 OK OK/S. Under

Thermo/Gem 62 OK OK

Man/Gem 56 Thick S. Over

Man/Man 53 OK OK pale

•Pattern consistent in all cases

•Pattern consistent for all users

•Thermo/Man best quality due to not batch banding

•Thermo/Gemini good, quicker, more consistent

SIZE M2

Per Unit TotalSIZE M2

Per Unit Total

MonoairHood

0.965x2

1.93 ClearvueH 500D 575W 645

0.37+hood

0.965

SensairHood

0.82x2

1.64 ThermotronH 1425D 715W 1095

0.78X2

1.56

Worktop 0.57x4

2.28 VaristainH 880D 710W 720

0.51+bench

1.12

Total 5.85 3.65

Manual MachineTime Walk away Time Walk away

Mon 3h30 1h 3h30 1h30

Tues 7h20 1h 3h20 1h55

Wed 6h25 1h 6h20 1h35

Thur 6h 1h 2h05 2h

Fri 4h55 1h 4h25 1h45

TIME 29h10 5h 19h40 8h45

TOTAL 24h10 10h55

PBS ml

Man Auto

BloodFluidOncology

299200390

380

Total

Litres

2.5 1.9

Cost £2.08 £1.57

•Gurr's

•Leishman's

•Trypsin

•Coverslip

•Mountant

Automatic method costs ~£80 more, per year

£• Saving: 689 MTO hours per year

• Extra cost: Machines + Extra Reagent + Maintenance

• Overall Saving• £6000 a year

QA 6 6Crossovers 8 5Space Needed 5.85m2 3.65m2

Time taken 4.72mins 2.11minsCost £1.76 £1.52

ConclusionStatistics on an average slide basis

Acknowledgements

• Eddy Maher

• Clive Dixon

• All at South East Scotland Cytogenetics Service.