SCS_Overview

Overview; iSaCS

For technicians and sales managers

This section describes the tasks, units and components of the 3DHISTECH Immunohistochemistry Staining automatic Cover slipping and Scanning equipment (iSaCS).

Features

· Complex and integrated solution with small footprint

· Handling of 72 slides simultaneously

· The modular construction allows to execute different tasks concurrently

· Immunohistochemistry and counterstaining of tissue specimen

· Application of stain during staining procedure

· Application of mounting medium during cover slipping

· Automatic scan of the created specimen in fluorescent (FL) or brightfield (BF) scan mode

· Automatic scan session performed in 30-120 seconds in brightfield scan mode for 1 specimen

Capacities

· 72 slides with specimen

· 150 cover slips

· 15 ml cover slip mounting medium (for 300 slides)

· 40 reagent vials

· 8 ml vial (for 80 staining procedures)

· 1 x 5l tank for washing fluid (for 200 slides)

· 1 x 5l tank for rinsing fluid (for 200 slides)

· 2 x 5l tanks for waste fluid (for 200 slides)

Main part list

Component		Stepper motor	Actuator	Sensor (ext.)	Light source	Camera
STA	Staining module	4	0	2	0	0
COV	Cover slipper	3	2	3	0	0
TRA	Transporter	3	1	1	0	0
WRU	Wash - rinse unit	4	2	12	0	0
REA	Reagent changer	1	0	0	1	1
SLI	Slide image	0	0	0	2	1
AIR	Air wiper	0	1	0	0	0
FRH	Frame; housing	0	0	4	1	0
MIDI	Digital scanner	6	0	2	3	2
Total	iSaCS	21	6	24	7	4

Units

The iSaCS is a complex, small footprint, integrated solution for the automation of tissue antigen marking. It is intended to perform IHC (Immunohistochemistry) staining, FISH (Fluorescent In Situ Hybridization) staining, counterstaining, cover slipping and scanning. The iSaCS was designed to use a modular construction approach, with the aim of having numerous modules working simultaneously.

Lumencor SPECTRA

The Lumencor SPECTRA Light Engine is an external light source that illuminates the field of view at fluorescent scanning procedure. It is connected to the iSaCS through the left side service door of the equipment with a light cable. The Lumencor has its own power cable through which it receives the necessary power.

Lumencor SPECTRA light engine®

Power tower

The main electronics of the iSaCS are stored in the power tower. It is located behind the scanner module in the equipment.

Power and Control

Slide storage

The user fills the slide rack (incubator) with slides containing the specimen.

For this purpose, 72 (12x6) slide bays are arranged in the rack.

To fulfill the tasks, dedicated slide bays are defined; always in these positions the named task will be executed; general purpose bays are used e.g. for sorting tasks.

· 1= Specimen Image; barcode capturing

· 7= Reagent application

· 11= Air wiper

· 13= Cover slipping

12 Wash and rinse chambers are available to stop the staining procedure (washing) and to remove washing fluid residues (rinsing).

Specimen image

· localization of tissue specimen on the slide

· slide identification through DataMatrix barcode label

· special illumination for tissue specimen with low contrast

Slide transporter

· fast positioning (approx. 5 s) of slides anywhere inside the slide storage and staining-cover-slipping unit

· dedicated motion schemes for different movement types

The transporter moves the slide from the slide rack position to

preview unit
staining unit
wash and rinse chambers
Air wiper position
cover slipping unit
any possible position
finally to the slide scanner and return to its origin position

Acoustic detector

To determine the quantity of required liquid exactly, an “acoustic fluid level detector” is implemented. The distance of the liquid surface to the pipette tip has to be known. If the distance is known exactly, the immersion depth of the pipette to gather the required liquid quantity for the process can be calculated, because the diameter of the vials is known.

· The distance of the liquid surface to the pipette tip is determined acoustically.

Exact gathering of quantity

It is important to gather the required quantity of the appropriate liquid very precise. At the end of the fluid application process, the pipette has to be empty! If the quantity of fluid in the pipette is too less, the process result becomes poor, otherwise, if fluid remains in the pipette, the level in the pipette will increase more and more and a waste of (often very expensive) fluid occurs. Furthermore, if the lfluid will dry in the pipette, waste process results are produced.

Reagent chain

· freely variable number (up to 40 pcs.) of 8 ml reagent vials

· vial identification through barcode Data Matrix labels

· disposable pipette tips (200 μl) dedicated to each reagent

· reagent level measurement with 100 μl precision

· reagent dispensing along any arbitrary trajectory

· precise covering of the specimen surface exclusively, using the lowest possible amount of reagent

Wash-rinse

· Washing and rinsing is done by the help of dedicated peristaltic pumps for distilled water, wash buffer and fluid waste.

· bubble detection in the tubing, priming of pumps with opto-tube sensors

· simultaneous use of multiple wash-and-rinse chambers possible, capacity: 12 slides

· precisely controlled flooding and draining of chambers, wash-and-rinse volume: 1-3 ml

· residual fluid amount on slide surface: less than 10 μl

Incubation

· small volume of single-slide incubation chambers minimizes evaporation

· incubation chambers in compact, small footprint arrangement, capacity: 72 slides

· control of relative humidity and temperature

Cover slipping

· Conventional glass cover slips of different sizes, capacity: 200 pcs.

· Verification of the number of available cover slips with +/- 1 pc. precision

· separation of multiple adhered cover slips

· aqueous mounting medium, capacity: 10 ml

· disposable pipette tip (200 μl) for mounting medium

· mounting medium level measurement with 100 μl precision

· pick-and-place cover slipping procedure using suction cup

· pressure monitoring in the vacuum system

· controlled and precise placement of cover-slip onto the sample

Workflow

General

The sample to be stained is placed on a glass slide together with its barcode and this will be inserted into the iSaCS.

Allowed slide dimensions are defined as:

Length: 75.00 to 76.00 mm

Width: 25.00 to 26.00 mm

Thickness: 00.95 to 01.05 mm

Specimen (sample) creation process

This process is done to create the specimen and finally, it will be placed onto the glass slide

· The creation of the specimen is done before the slide will be inserted into the slide rack of the iSaSC.

More, detailed information about the sample creation process can be found in

Introduction to specimen preparation and

An animated guide to immunohistochemistry (IHS)

Place slides

· Remove slide container

Place slides

The user can fill slides in any bay of the slide rack; a special sequence is not required.

· Insert the slide container

Start process

While the process starts, the slide mover senses 3 slide bays for the presence of the slide by using a special algorithm. If a slide is found, this will be moved to the slide image unit.

If the image of the specimen (and barcode capturing of the slide) is made, the slide will be returned to its origin place.

Slide image

The unit makes an image of the

· slide’s scan area and the

· Barcode area.

This way, the position and the size of the sample is recognized and the staining procedure will be executed with the entire tissue. On the other side, because the staining materials are very expensive, redundant areas of the slide will not be coated with staining fluid.

The barcode of the specimen contains information about the specimen creation process, dyes, staining and about the scan process also; how to handle the specimen

The user can define the

· Number of different dyes for the sample

· Type of each dye and sequence

· quantity (area) of the appropriate dye

· residence time of the dye

Staining

After the residence time of the appropriate dye is passed, the unnecessary dye will be removed from the sample by washing and rinsing it and drying it up.

If the incubation time is over, the staining with the next dye follows or, if the staining process is finished, the slide moves to the cover slipping unit.

Reagent code

Before the reagent vial is placed into the vial chain, the reagent code, containing the name, staining time and other process relevant parameters has to be affixed onto the bottom of the vial.

The software, during the startup procedure reads the code of each vial and stores process relevant information. Tis way, the position in the chain is defined also.

Before the staining procedure starts, the required vial is moved to the vial using position and the pipette tip immerges into the reagent and gathers the required quantity.

The sample will be stained by dropping dye onto it.

Washing

Rinsing

Cover slipping

If the rinsing and wiping process of the specimen is finished, the glass slip will cover the specimen.

With a pipette paraffin will be spend to cover the specimen and by fitting the cover slip, the paraffin works as an adhesive bonding mean.

Scanning

After the specimen creation process the content of the slide will be scanned.

The scanning procedure in the 3DHISTECH iSaCS is as follows:

· The slides are inserted into the scanner and removed automatically via a slide holder tray.

· The slide bays in the slide holder tray allowing an autonomic work of the scanner.

· Designed for brightfield scan sessions and fluorescent scan sessions.

· The scanner part is based on a modular MIDI scanner and creates the virtual specimen

· An automatic scan session is performed in 30 … 60 seconds for 1 specimen in BF scan mode.