KOS is a microwave Multifunctional Tissue Processor that fits
on a countertop. It is able to perform Tissue processing for biopsy samples up to 3mm in
size, Decalcification procedures, Special stains, Fixation, Gross hardening and Antigen
retrieval.
This state of the art processor is ideally suited as a stand-alone tissue processor or
to complement existing processing capabilities found in histology laboratories where
multitasking nature of the instrument can enhance and accelerate many of the typical
applications encountered.
KOS is smart, icon driven software makes this intuitive and easy to use.
KOS is fast. Factory prestored optimized fully documented protocols for rapid microwave
enhanced procedures. No calibrations are required. Just plug in and process.
Reliable temperature sensors for the protection of your precious specimens.
Flexible user defined customized programs can be created and added to a most used
favorites list for quick access as required for meeting the needs of each user in the
laboratory. The Kos Rapid tissue processor will enable the laboratory manager to fulfill
the requirements set by the C.L.S.I. and CAP inspection guidelines, it is an ideal
substitute for the “Kitchen Type” microwave utilized in many histology
laboratories.
Rapid tissue processing is the primary application requested by most KOS users. The
compact footprint makes it ideal for small laboratories, the countertop design puts the
unit at comfortable working level. The easy Icon driven menu allows for ease of use and
consistent reproducible results. The KOS concept includes a Microwave tissue processor,
specially developed Histomodules and a set of pre-stored programs. Three configurations
are available to meet the needs of any laboratory. Small batches of 1 to 4 cassettes
utilize the 4K module this allows flexibility for STAT processing of transplant or
urgent biopsies in as little as thirty minutes. Process up to 45 cassette batches using
the 45K module or increase capacity with the new 110 Cassette configuration. Kos uses
standardized protocols developed and validated over 20 years of user application. The
easy icon driven menu is intuitive, the user simply follows the prompts to move from
fixation through dehydration, clearing and wax infiltration. Kos is semi-automatic and
requires minimal interaction from the user. Hands on time is less than 10 minutes per
batch.
The Milestone protocols are safe, no xylene is required. While Proprietary reagents are
available the KOS is an open system using Ethanol and Isopropanol protocols for all
types of specimens (up to 3mm thick) to produce high quality slides. There is no
compromise in regard to morphology and staining expression. Staining properties of
microwave processed specimens are excellent for routine H&E staining as well as HC,
IHC and FISH.
The KOS provides state-of-art microwave Hardware/Software, for obtaining consistent and
reproducible results in the histopathology laboratory, for high throughput tissue
processing and other histological applications. The system and its parts have been
engineered for laboratory use and can therefore withstand the harshest laboratory
conditions for continuous use in Histological sample preparation procedures.
Hardware and safety features: The KOS has dedicated software, based on Windows CE
(Microsoft Windows CE is a trademark of Microsoft Corporation). Use the touch screen pen
to select the desired menu function. The KOS is neither a kitchen type nor a
commercially modified kitchen type microwave unit. It is a rugged, heavy duty
professional system, designed to guarantee many years of trouble free and safe
operation.
No need to waste precious hood space the KOS is self-vented and self-contained. The
exhaust fan with built-in rechargeable charcoal filter eliminates toxic vapors from the
laboratory. KOS may be connected to an existing exhaust system if desired.
Auto start magnetic stirrer provides homogenous temperature distribution within the
solutions, eliminating hot spots. All samples are processed under the same preset
condition. Infra-Red sensor provides fully automatic noncontact temperature control.
This allows for 100% walk-away operation.
Histoprocessing: Preset protocols for specimens 1mm, 2mm and 3mm in thickness are
preprogramed for ease of use and consistent performance. Ethanol-Isopropanol-wax and
Prowave (a single step Milestone proprietary reagent) options are available in the menu.
User selects the desired option, enters the tissue thickness, and module. A program is
then created. The user has the choice to include a heated fixation step in 10% formalin
for FIneFIX as needed and desired. The KOS will prompt the user to perform two alcohol
rinses after the formalin to eliminate water carryover into the single dehydration
phase. The single Ethanol dehydration phase is heated to 65C. At the conclusion of the
Ethanol dehydration the rack is transferred to Isopropyl alcohol for the clearing phase.
Isopropyl provides gentle and effective clearing without the hazards associated with
xylene. The Isopropyl step is heated to 68C. The tissue is finally moved to a preheated
Paraffin module for paraffin infiltration.
Typical processing times
Transplant =30 minutes including fixation using Prowave single solution protocol
1mm 45K module= 1 hour 10 minutes plus 15 minutes to include heated fixation
2mm 45K module =2 hours 15 minutes plus 20 minutes to include heated fixation
3mm 45K module =3 hours and 5 minutes plus 30 minutes to include heated fixation
Bone Marrow Processing. Same day diagnosis for Bone Marrow trephine biopsies. Kos is
able to provide Decalcification and processing in less than three hours. The ideal
application of the multifunctionality of the KOS. Optimized decalcification with your
choice of EDTA or Formic Acid followed by xylene free rapid processing protocol. This
provides the ability to offer accurate diagnostics and fast turnaround times.
Typical Bone marrow processing times using 4K module. Up to 4 cassettes.
Fixation in 10% Formalin 50C for 10 minutes. Rinse in tap water 10 minutes.
Decalcification with Formic Acid 0 minutes at 50C. Rinsing in Ethanol 2 minutes. Tissue
processing with Prowave 15 minutes, Impregnation in wax 23 minutes. Total time 80
minutes.
An excellent application for KOS is using it to perform rapid Bone Decalcification.
Decal bone in hours not days.
There are 2 types of decalcifying agents: ACIDS and CHELATING AGENTS.
ACIDS: they can be strong and inorganic: most commonly used are NITRIC <10% and
HYDROCHLORIDRIC
<10%, or weak and organic: most commonly used are FORMIC <20% and ACETIC
<10%.
In the case of particularly strong acids, the specimens must be properly fixed to slow
down the digestion of
tissue components, decalcification must be monitored carefully to avoid over
decalcification in which nuclear detail is lost,
and antigenic sites damaged.
CHELATING AGENTS: bind calcium slowly and gently by forming an insoluble non-ionized
complex, so it is the
preferred method in case of need of subsequent analysis like electron microscopy,
immunohistochemistry and
molecular biology. EDTA (Ethylendiaminetetraacetic acid) is the most common chelating
agent.
The rate at which EDTA will decalcify is pH dependent. The optimal pH is 7.0-7.4.Because
the chelating process is very slow, this reagent is not suitable for urgent LARGE and
HARD specimens
but more appropriate for BONE MARROW and SOFT LITTLE BONES, where very high quality
morphology is
required or particular molecular elements must be preserved for techniques such as IHC,
FISH or PCR.
WHAT IS MILESTONE CHELATING AGENT “MOLDECAL”?
MOLDecal is an innovative decalcifying agent for optimized molecular results.
It consists of a proprietary mixture of EDTA salts to obtain an ideal decalcification pH
of 7.2-7.4, with no
addition of buffer. When combined with agitation and temperature control the MOLDecal
can reduce the
decalcification time while maintaining a high recovery level for nucleic acids.
WHAT ARE THE FUTURE DIRECTIONS FOR DECALCIFICATION?
Molecular diagnostics are rapidly becoming part of the standard of care for cancer
patients. This biomarker
based approach serves to select patients likely to benefit from targeted treatment
regimens and potentially
assists in their inclusion into appropriate clinical trials.
Bone is a common and occasionally the sole site of metastasis or sampling. Bone
biopsies, however present an
unique challenge to molecular diagnostics, as they are subject to decalcification during
routine histology
processing. Unfortunately, commonly used decalcification agents containing strong acids
(i.e. HCL, Nitric acid)
result in degradation of nucleic acid required for subsequent molecular profiling and
biomarker analysis.
WHICH FACTORS INFLUENCE DECALCIFICATION?
1. Heating – It accelerates the chemical reaction.
2. Agitation – It avoids the saturation of the solution with “fresh” reagent in
contact with the surface of the
specimen at all times.
3. Size of sample – The larger the specimen the longer it will take to achieve
decalcification.
4. Type of tissue – Compact/cortical bone is hard, solid and strong while
spongy/trabeculae bone is soft and
easier to decalcify.
5. Type and concentration of decalcifying agent.
Q. HOW TO CHOOSE A DECALCIFYING AGENT?
In most laboratories buffered EDTA is used especially for decalcification of bone
marrow
With a decalcification time of 2/3 hours.
By checking the MSDS it is clear that decalcification is carried out by the buffer which
is a
Strong mineral acid, in most cases 10%HCl.
WHAT ARE THE FUTURE DIRECTIONS FOR DECALCIFICATION?
Molecular diagnostics are rapidly becoming part of the standard of care for cancer
patients. This biomarker
Based approach serves to select patients likely to benefit from targeted treatment
regimens and potentially
Assists in their inclusion into appropriate clinical trials.
Bone is a common and occasionally the sole site of metastasis or sampling. Bone
biopsies, however present an
Unique challenge to molecular diagnostics, as they are subject to decalcification during
routine histology
Processing. Unfortunately, commonly used decalcification agents containing strong acids
(i.e. HCL, Nitric acid)
Result in degradation of nucleic acid required for subsequent molecular profiling and
biomarker analysis.
Q. WHICH FACTORS INFLUENCE DECALCIFICATION?
1. Heating – It accelerates the chemical reaction.
2. Agitation – It avoids the saturation of the solution with “fresh” reagent in
contact with the surface of the
Specimen at all times.
3. Size of sample – The larger the specimen the longer it will take to achieve
decalcification.
4. Type of tissue – Compact/cortical bone is hard, solid and strong while
spongy/trabeculae bone is soft and
Easier to decalcify.
5. Type and concentration of decalcifying agent. The efficient stirring continuously
keeps the bone surface in contact with “fresh” decalcifier, positively influencing the
equilibrium. The use of gentle heat accelerates action of the decalcifying solution.
EDTA is the preferred solution for molecular studies but can take days or even weeks to
decal large or dense bone samples with EDT. Utilizing the KOS Decalcification time in
EDTA can be reduced dividing your routine decalcification time by 2.5 if you wish to use
37C. At 50C the time for decalcification can be divided by 5.
For the first run the decalcification time setting can be roughly calculated by dividing
your routine
Decalcification time by 2.5 at 37°C, or by 5 at 50°C, .i.e.: routine room temperature
decalcification time 25 hours; KOS set time: 10 hours at 37°C.i.e.: routine room
temperature decalcification time 25 hours; KOS set time: 5 hours at 50°C.
If the decalcifying reagent is a WEAK ACID LIKE FORMIC ACID 10-12%:
For the first run the decalcification time setting can be roughly calculated by dividing
your routine
Decalcification time by 10 at 37°C, by 5 at 50°C.
i.e.: routine room temperature decalcification time 8 hours; KOS set time: 50 minutes at
37°C.
We strongly advise to checking the specimens every 30-60 minutes.
In case faster turnaround time is needed, it is possible to test at 50°C but in this
case we strongly advise
Checking the specimens every 30 minutes i.e.: routine room temperature decalcification
time 8 hours; KOS set time: 25 minutes at 50°C.
Hydrochloric Acid decal solutions or RDO should not be used in KOS.
There are several rack options available for bone decalcification with KOS. The 4K
module may be used for 104 cassettes. Up to 45 Cassettes or 110 Cassettes rack are
available as well as super Mega and Super Mega slim racks. Typically once decalcified
larger samples over 3mm in thickness are processed using LOGOS, DELTA or Magnus or
placed on a conventional processor.
Special Stains can be performed up to 76% faster when using KOS for histochemical
staining protocols several time consuming staining procedures are enhanced by microwave
irradiation. KOS has factory stored, optimized, step by step protocols for the most
common stains for 1-40 slides. Use the preset protocols or customize to guarantee
operator independent, consistent results. The specialized modules include a stirrer that
that allows for temperature uniformity along the entire length of the slide.
The following special stains are included in the KOS menu.
GMS for Fungi, Grimelius (Argryrophil), Masson Trichrome, Fontana Masson, PAS, Alcian
Blue, Ziehl Nielsen, Elastica (Orcein) Pearls Iron Stain.
Reporting. At the conclusion of every process the user has the ability to save a report.
This report documents user, time started, protocol in use, time and temperature of each
phase. It is saved in the memory and may be downloaded via USB to be reviewed or saved.
Each KOS comes with an electronic user manual and copy of Logviewer software to allow
for review and printing of reports from a PC.
Reagent management options. The user may desire to set up reagent management parameters
to track and signal expired reagents. These are cassette count based.Kos may be used to
perform Time at Temperature or Antigen retrieval procedures for IHC. Simply select the
temperature, select the histomodule, select time and press go.All protocols used
routinely may be entered into a special Favorites menu. This is easy to do simply press
and hold the small heart on the protocol screen. This allows users to easily access
protocols in frequent use, standardization is simplified as the correct program is
always at your fingertips.