Immunohistochemical staining automatical coverslipping ......Annamaria Csizmadia, 3DHISTECH Ltd....
Transcript of Immunohistochemical staining automatical coverslipping ......Annamaria Csizmadia, 3DHISTECH Ltd....
Immunohistochemical staining
automatical coverslipping scanner
iSaCS
Annamaria Csizmadia, 3DHISTECH Ltd.
Dig
ital
imag
es,
Macrosta
tion
Au
tom
ati
cla
bo
rato
ry
instr
um
en
t
Tissue/Sample fixation
Grossing
Embedding
Deparaffination
Sectioning
Antigen retrieval
IHC staining
Coverslipping
Scanning
Image Analysis
Digital methods in the histology workflow
Scanner
Stainer
Staining-Coverslipping-Scanning Unit
Pannoramic MIDI
Leica BOND MAX
Agilent technologies, DAKO Coverslipper
Coverslipper
Leica Bond Max
Dako Omnis VentanaBenchMark
ULTRA
iSaCS
Deparaffination + + + -
HIER + + + -
Blocking of endogenoous enzyme + + + +
Primary antobody + + + +
Secondery antibody + + + +
Chromogen substrate + + + +
Counterstain + + + +
Mounting - - - +
Coverslipping - - - +
Scanning - - - +
Image Analyis - - - +
Automated phase of different stainers
Leica Bond Max Dako Omnis Ventana BenchMark ULTRA iSaCS
Slide capacity 30(10 slides/rack)
45(5slides/rack)
30(Individual chamber)
36(72 slides/rack)
Runtime 3:30 2:30 2:30 5:45
Slides/day 90 165 90 108
Number of Reagents
36 36 35 40
Contonous staining yes yes yes no
Staining Principles Capillary Gap Staining Dynamic GapStaining
Liquid Overly Technology Open individualslide staining
Deparaffination;HIER
yes yes yes no
UsedAntibody/slide
100µl 100µl 100µl 47µl
First digital slide Min. 04:00 Min. 03:00 Min. 03:00 02:15
Comparison of different stainers
iSaCS - System architecture
1. Digital slide scanner
2. Slide preview module
3. Slide moving arm
4. Pipetting module
5. Reagent changer
6. Incubation module
7. Wash & rinse chamber
8. Air wiper module
9. Coverslipping module
Scanner, Preview imaging, Slide moving arm
1. Digital slide scanner:
•Improved Midi scanner
•Flash scanning technology
•Fluorescent modification
2. Preview imaging of unstained samples:
•Special high-contrast illumination
•Localization of tissue sample on the slide
•Reagent amount based on tissue size (µl/mm2)
•Dispensing trajectory covers tissue area exclusively
•Slide identification by 2D code
3.Slide moving arm:
•Transports slides between modules
Pipetting unit, Reagent changer
4. Pipetting unit:
•Liquid level sensor
5. Reagent changer:
•Reagent carrier chain: adjustable number of chain links (max. 40)
•Chain link: 8 ml reagent vial + disposable pipetting tip (200 µl)
•Reagent vial identification by 2D code, pipetting tip sensor
•Reagent tray with lid: removable, fits into the refrigerator
Coverslipping unit, Incubation tray
8. Coverslipping unit:
•Mounting medium (aqueous, aqueous with hardening)
•Disposable pipetting tip (200 µl)
•Glass coverslips (22x50 mm)
•Vacuum: pump,
•Pick-and-place suction cup
•Separation of coverslips
•Precise positioning eliminates bubbles
•Liquid level sensor
•Pipetting tip sensor
9. Incubation tray:
•72 slides in compact arrangement
•Chambers eliminate evaporation
Wash & Rinse unit, Airwiper
6. Wash & Rinse unit:
•11 parallel Wash chambers
•1 chamber only for rinsing
•Flooding/draining
•3 peristaltic pumps:
• Wash buffer, DI water
• Hazardous / Non-H waste
•4 removable 5l tanks: tank ID
•Tank cap: tubing, level sensor
7. Airwiper removes liquid film:
• Airblade avoids desiccation
• Suction tray avoids spilling
Prepared slide forstaining
PIS
/LIS
Recues
sta
inin
gpro
cedure
Slide recognation
Slide Barcodereading
ID recognationbased on
Label Area
Staining arearecognation based on
DFP
Protocolcreation
ProtocolAssignement
Extrenal request from Database User-defined System-defined
PREPARATION
STAINING
Staining of the definedtissue area
COVERING
Coverslipping of the stained
slide
IMAGE ANALYSIS
SCANNING
Scanning of the stained
area
Image analysis of whole slide
Trecking/Database
iSaCS sample detection
Active area for stain
iSaCS Protocol Editor
Structure of Protocol
Protocol
Protocol parts
Blocking Primery antibody Secondary antibody Labeling Finishing
Pro
toco
lste
ps
- Pipetting- Incubating
- Rinsing- Washing
- Covering- Scanning- Quantification
Reagent recognition
Undetermined vials
- The system recognized them as empty vials
- Manual Reagent attachment
Automatic reagent recognition
- Predefined reagent type to the vial
- Deliver with the system
Changes in the software of iSaCS
New failure management :
- Covering unit: we check the vacuum after the coverslip is taken up
- Scan : Continous communication between the staining and
scanning parts
Improuved reagent recognation:
Virtual reagent card shows the needed reagent type for the staining
Emergency STOP
Changes on the hardware
Covering unit:
- Opto sensor for X axis
- Keyence Sensor for tip detection
- Mounting medium level engraving
- Mounting medium level clamp
- Accessories gate
Moving arm:
• Belt guide
• Fixed vane
Changes on the hardware
• Pipetting unit:
- Opto sensor for X axis
• Accessories:
- Teflon coated reagent chamber
- Complete new tanks
- Black painted incubator
- Slide deflector for the incubator chamber
iSaCS – The main features
A single device integrating an immunostainer, a coverslipper and a scanner
Cost-effective open system, large capacity, bench-top construction saves lab-space
The patented automatic sample detection makes only the effective sample area
gets stained
Standardized and controlled sample treatment without human intervention
Parallel task execution, modular construction
First stained slides in CaseCenter can be available on-line right away:
speedy evaluation
iSaCS – The main features
Digital scanning and staining optimized using pre-staining preview images
Fluorescent IHC features: preview, scanning, archiving without degradation
Quick protocol definition, and easy handling of consumables
iSaCS scanning device can be used separately as individual scanner
Image analysis process will be run automatically on the whole slide
Slide and protocol recognition based on barcode
Easy to use software with hierarchical user interface
What will be new in 2018?
• New integrated scanner workflow into iSaCS software
• Operation log under the staining/covering/scanning process
• Auto Dilatation on the preview of tissue
Thank you for the attention