1. FISH Automation 소개
진단검사의학과
강지연

2. Fluorescence in situ Hybridization
검사하고자 하는 표본에 찾고자 하는 probe를 반응시킨 후 특정부위의 유전자나 염색체의 결실, 추가, 증폭, 재배열을 형광현미경으로 관찰하는 방법
Specimen treatment
Harvest / Slide preparation 및 경검
Front End FISH
1st day
2nd day (암실에서 진행)
Pretreatment
Dehydration
암실에서 진행
DNA-Probe 첨가
Denaturation
Hybridization
Cover slip 제거
Washing (Wash1,Wash2,Wash3)
Counter stain: DAPI 부착

3. Back End FISH Analysis by fluorescence microscopy
dual color, dual fusion
break apart

4. FISH Automation - Overview
FISH Automation - Overview
Front End Automation
Automates slide/specimen pretreatment and assay
Can be used for FISH, G-Banding, IHC & others
Two Configurations
(1) VP2000 and ThermoBrite™
(2) Xmatrx
Back End Automation
Automated slide scanning & enumeration
MetaSystems
Other Applications in process
We sell and support FISH however the instruments can be used for other slide-based techniques.
VP2000 and ThermoBrite can be used separately and do not have to be used in succession

5. Automation Choices Front-End Back-End VP2000 ThermoBrite metasystems
Denaturation
&
Hybridization
ThermoBrite
Automation Choices
Slide
Pre-treatment
& Washing
VP2000
Slide Scanning
& Enumeration
Slide
preparation
Pre-treatment,
Washing,
Denaturation,
Hybridization &
Final Cover slip
Xmatrx*
metasystems
* In Development

6. Front End FISH VP2000 Update
Automates FISH Assay Steps:
FISH Pretreatment
Specimen Staining
Up to 50 slides simultaneously
3 heated reagent basins
Ventilated closed system
General purpose system accommodates most slide-based assays: histology, G-banding, etc
Increase FISH throughput
Increase consistency of results
Decrease labor costs

7. Front-End FISH ThermoBrite™
Front-End FISH ThermoBrite™
Slide Hybridization &
Denaturation System
Eliminates separate denaturation
40 definable programs – LCD screen
12 slide capacity – guides hold slides
Humidity cards
Superior temperature uniformity
Rapid temperature ramp

8. Front-End FISH XMATRX Automation of ALL FISH assay steps Throughput
Front-End FISH XMATRX
Automation of ALL FISH assay steps
Pre-treatment, Probe, Coverslip application, Washing, Denaturation, Hybridization, Cover Slip Removal, Wash, Counter stain & Final Cover slip
Throughput
Up to 40 slides per run
Up to 80 assays per run
Random access, up to 8 different assays in one run
Continuous access, slides can be added or removed during the run
Full reagent management capability
Applies, removes & reapplies cover slips
PC with easy to use software

9. Front-End FISH A Fully Automated FISH System

10. Back-End FISH Carl Zeiss Microscope
A Team Meeting Marketintroduction LSM 5 PASCAL, LSM 510 SW Rel. 2.5, ConfoCor 2 Stand Alone
Back-End FISH Carl Zeiss Microscope
Axio Imager.M1/Z1
Motorized stand
Automatic Focussing
Maximum variability
Control by PC or touch screen
The best fluorescence
with higher contrast 1

11. Metafer - Slide Scanning System
A Team Meeting Marketintroduction LSM 5 PASCAL, LSM 510 SW Rel. 2.5, ConfoCor 2 Stand Alone
Back-End FISH
Metafer - Slide Scanning System 1

12. real-time image analysis and microscope control
A Team Meeting Marketintroduction LSM 5 PASCAL, LSM 510 SW Rel. 2.5, ConfoCor 2 Stand Alone
Metafer
Concept
CCD Camera
Metafer
PC Win 9x 2000 / NT
Motorized stage (x,y)
real-time image analysis and microscope control
Motorized microscope (z) 1

13. A Team Meeting Marketintroduction LSM 5 PASCAL, LSM 510 SW Rel. 2
A Team Meeting Marketintroduction LSM 5 PASCAL, LSM 510 SW Rel. 2.5, ConfoCor 2 Stand Alone
Metafer
Concept
• Microscope slide is scanned field by field at lowest possible
magnification without gaps
• Objects of interest are automatically identified ; gallery images,
object features and positions are recorded 1

14. Metafer Advantages of Metafer High speed Slide Scanning (70mm/s)
A Team Meeting Marketintroduction LSM 5 PASCAL, LSM 510 SW Rel. 2.5, ConfoCor 2 Stand Alone
Metafer
Advantages of Metafer
High speed Slide Scanning (70mm/s)
High quality image capture during the scan
High throughput and capacity
multi-fluorochrome capability from UV to IR
- very few objects available for analysis
- statistical analysis of many objects 1

15. Metafer Applications MSearch : metaphase finder
A Team Meeting Marketintroduction LSM 5 PASCAL, LSM 510 SW Rel. 2.5, ConfoCor 2 Stand Alone
Metafer
Applications
MSearch : metaphase finder
- Micronucleus analysis
- Automatic analysis of dicentrics
MetaCyte : automated FISH analysis / spot counting
- spot counting and translocation analysis
- amplification analysis in tissue section
- Comet scan
- High content screening
RCDetect : rare cell detection
- minimum residual disease detection 1

16. Metafer - MSearch : metaphase finder
A Team Meeting Marketintroduction LSM 5 PASCAL, LSM 510 SW Rel. 2.5, ConfoCor 2 Stand Alone
Metafer - MSearch : metaphase finder 1

17. Metafer – Msearch : metaphase finder
A Team Meeting Marketintroduction LSM 5 PASCAL, LSM 510 SW Rel. 2.5, ConfoCor 2 Stand Alone
Metafer – Msearch : metaphase finder
Fast metaphase finding in transmitted and fluorescent light 1

18. Metafer - MetaCyte : automated FISH analysis
A Team Meeting Marketintroduction LSM 5 PASCAL, LSM 510 SW Rel. 2.5, ConfoCor 2 Stand Alone
Metafer - MetaCyte : automated FISH analysis 1

19. A Team Meeting Marketintroduction LSM 5 PASCAL, LSM 510 SW Rel. 2
A Team Meeting Marketintroduction LSM 5 PASCAL, LSM 510 SW Rel. 2.5, ConfoCor 2 Stand Alone
Metafer – Slide Feeder 1

20. Isis - Fluoresc. Imaging System
A Team Meeting Marketintroduction LSM 5 PASCAL, LSM 510 SW Rel. 2.5, ConfoCor 2 Stand Alone
Back-End FISH
Isis - Fluoresc. Imaging System 1

21. Isis Fluoresc. Imaging System – features and benefits
Fast and easy capture
Powerful image enhancement
Various color display mode, measurement function
Database manegement as Ikaros
Additional Isis modules
Color karyotyping module
mFISH and mBAND module
CGH(Comparative Genome Hybridization) module
Telomere measurement module

22. Isis Fluoresc. Imaging System – Capture
A Team Meeting Marketintroduction LSM 5 PASCAL, LSM 510 SW Rel. 2.5, ConfoCor 2 Stand Alone
Isis Fluoresc. Imaging System – Capture
Focus adjustment live on screen
Automatic exposure control
If with Carl Zeiss Axio Imager.M1
or Z1 microscope
- automatic filter change
- focus stack capture 1

23. Summary
Improve workflow efficiency
Increase throughput; enable labs to do more with the same resources
Reduce “hands-on” time; free up resources and simplifies workflow
Increase assay quality and reproducibility
Multiple solutions; different levels of automation
Applications to our laboratory(metasystems)
Rapid finding of metaphase & FISH interphase
Good assay quality of FISH analysis
Pannel FISH
Urovision