Operation Manual

MTC Cryostat

SLEE MEDICAL GMBH MAINZ

Lise-Meitner-Str. 1
55129 Mainz / Germany
FON 49/6131/95871-0
FAX 49/6131/95871-22
Email: email@sleemainz.com
Internet: www.sleemainz.com

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LIST OF CONTENT

INTRODUCTION 3
Important Information 4
Safety Aspects 5
Description of Unit 7
Accessories 7

ARRIVAL AND INSTALLATION 8

Receipt 8
Unpacking 8
Installation 8
Connection to Main Supply 8

START UP OF UNIT 9
Installing Microtome 9
Fitting and checking operation elements 9
Controller 12

PRDUCING SECTIONS 14
Microtome knife (cutting edge, knife angle) 14
Relief or clearance angle 15
Knife holder 15
Adjusting and fixing of knife 15
Anti-Roll Plate 16
Trimming 17
Cutting 17

OPTION: DISPOSABLE BLADE HOLDER 18

GUIDE FOR CUTTING TEMPERATURES 19

TROUBLE SHOOTING 20

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INTRODUCTION

Dear Customer,

You have acquired a MTC cryostat of the SLEE Medical GmbH meeting highest quality
standards. Please carefully read these constructions before starting to operate the unit.

The use of unfixed material in histology demands highest requirements regarding quantity and
resulting quality from the user and the unit. Your cryostat meets these demands

The operating instructions serve as a guide and support for the start-up and routine use of the
freezing microtome for beginners or experienced users. Please contact us if you require
additional information.

The unit is primarily designed for the following applications:

• Routine histology
• Fast biopsy diagnostic
• Fast sectioning in prosector
• Immune histo-chemistry
• Autoradiography
• Enzyme histo chemistry
• Fluorescence microscopy
• Analytical histo-chemistry
• Micro-chemistry

As a portable bench model its application is versatile for experimental, diagnostically, analytical
and scientific research.

Your recommendations and questions are welcome at all times.

SLEE MEDICAL GMBH MAINZ

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IMPORTANT INFORMATION

This manual includes important information about the operation and safety aspects of the unit.
The manual is an important part of the unit. It must be read by the user and should always be
kept near the unit.

Please read in advance the details concerning the safety aspects even if you know cryostates or
other SLEE products already.

In case there is the necessity for further requirements regarding the operation of cryostates in
some countries, these have to be taken into account and these instructions should be kept
together with this operation manual.

SLEE cryostates should only be used by trained personell.

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SAFETY ASPECTS

SLEE cryostates are provided with the following safety equipment:

- Hand wheel lock

- Stanndard knife- and disposable blade holder provide with finger protection

The institution which owns the unit and the persons working with the unit, servicing or repairing it
have the responsibility for a hazard-free use.

Hand wheel stop

Always use the finger protection with the standard knife-/disposable blade holder and put hand
wheel in stop position,
- Before working with the knife or specimen
- Before changing the specimen
- During break time

The hand wheel can be stopped in the upper position. For this purpose move the lever towards
the red point, it will engage in the upper position.

For releasing the stop please move the knob towards the green point.

Finger protection:
Use the finger protection always
- Before you start any work with the knife or specimen
- Before changing the specimen
- During work break

Electrical power connection:

Do not use any extension lead. Make sure that electric power is constant:
- This should be examined during installation of the unit by a competent person
- Use a dedicated fuse for the unit
- Do not connect another unit to the same power circuit
- Check if the voltage of the mains supply is identical with the name plate of the unit

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Working with microtome knives and disposable blades

Microtome knives and disposable blades have extremely sharp edges and this can lead to
injuries.

Please be extremely careful when handling microtome knives and disposable blades. Do not
place microtome knives or disposable blades at unsecured areas. Never position microtome
knives or disposable blades with the sharp edge pointing upwards.Never try to catch a falling
microtome knife. Always insert the specimen first and then the microtome knife or the disposable
blade.

Cleaning of the unit

For cleaning the unit only use alcoholic media, not acetone or xylene. Never spray or use
cleaning medium directly onto operating foils. Please take note of the safety aspects of the
manufacturer. Please use gloves.

Service:

Besides regular cleaning no service is necessary for the unit through the user. SLEE suggests to
do a ½ to 1 year regular service through a SLEE authorized service technician. Internal
components should only be serviced by technicians authorized by SLEE.

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DESCRIPTION OF UNIT

The freezing microtome comprises three assembly groups:

• Body with cooling chamber and refrigeration unit

• Electronic control for best cooling results
• SLEE MTC-microtome with exterior operating chamber

The body is powder coated sheet metal. It encloses the cooling chamber. The isolation is energy
conserving. The refrigeration unit is in the rear of the freezing microtome. The cooling medium is
CFC-free.

The technical parameters are integrated and stored in the control unit.

The unit is provided with automatic defrost commencing at midnight. Defrosting removes
condensed water from the evaporator. For automatic defrost adjustment of actual time is
important.

The MTC-microtome has been especially manufactured for use at low temperatures and
guarantees very good sectioning quality at these temperatures. All exposed materials are made
of stainless steel. All microtomes have been individually tested.

ACCESSORIES

1 Microtome knife C 120 mm with wooden box

2 Specimen holders
1 Set of tools
1 Embedding medium GSV 1
1 Cryo-oil
1 Brush
2 Finger protection

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ARRIVAL AND INSTALLATION

2.1 Receipt

The unit has been thoroughly tested under operating conditions and following stringent quality
control procedures. For transport the microtome and operating segments may have been
disassembled and these items may have to be fitted again. If unit is started up later, ensure that it
is stored dry and protected against contamination.

Examine if there has been transport damage and inform manufacturer accordingly.

2.2 Unpacking

Remove packing material and compare delivery note with delivered items. Microtome, operating
elements and accessories are in the cryostat chamber. Remove packing material only when
necessary, so that quality does not suffer.

2.3 Installation

The freezing microtome weighs 60 kilos. Select a suitable work position for upright installation.
Direct solar radiation should be avoided. Placing unit in air-conditioned rooms or in direct vicinity
of exhaust systems may influence the cooling circulation of the unit unfavourably.

2.4 Connection to Mains Supply

Compare voltage on nameplate with available voltage. Operate only if voltage is the same.
Phase acquaint us if this does not agree since conversion is no problem.
The connection of unit to mains is via main cable. In addition at the rear of the unit there are
fuses for unit protection.

Please acquaint yourself with programming instructions before start-up.

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START-UP OF UNIT

3.1 Installing microtome

The MTC microtome is a rotary microtome and may frequently be disassembled for transport
reasons. In this case it needs to be installed into the cryostat.
To do this turn the hand wheel on right-hand side so that the coughing has an upward incline of
20° inside the chamber. Carefully place microtome with specimen holder so that coupling counter
part aligns with coupling. Push couplings into each other and observe at the same time that guide
rods on the left and right grip into the guide. Then the microtome is fixed by means of hexagon
key (Set of tools) at the front. For proper start-up it is necessary that the beyer of the hand wheel
and specimen holder are in top position simultaneously and that the microtome gives a well-
balanced impression when turning handle. This means that the microtome runs easily and
smoothly in any position.

3.2 Fitting and checking Operation elements

Operation elements comprise right-hand and left-hand hand wheel, specimen clamp, specimen
orientation and micrometer adjustment.
If microtome is not installed, first proceed as per paragraph 3. 1.
The left hand wheel, in the following called coarse advance, is for the horizontal movement of
knife to specimen or away from specimen. One revolution is 100 microns. The knob above the
hand wheel is part of the anti-roll device and moves the anti-roll plate.
The coarse advance shaft must be connected to microtome. The shaft is pushed into bushing
and is screwed with hexagon key. The fiat surface of the shaft is receiving point of screw. Insert
the dirt collection sheets. Then push anti-roll shaft into bushing and screw with hexagon key.
Again the fiat surface of the shaft allows fixing screw.

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If hand wheel handle is in highest position, the specimen too should have highest position. By
turning hand wheel up- and down movement of specimen is obtained. The unit is balanced by
flyweight to allow smooth turning of hand wheel. If this is not the case, inform service since
adjustment of flyweight may be necessary (see service manual). For safety reasons the hand
wheel should be locked in top position when manipulating at specimen holder.

The right hand wheel, in the following called cutting wheel, is for movement of specimen in
relation to knife. Specimen orientation is for receiving specimen holder and for horizontal and
vertical positioning in relation to knife.

By tightening or loosening the right-hand Lever the specimen holder is fitted or removed. The heft
hand Lever is for positioning and fixing specimen holder in desired position.

The micrometer adjustment is at the front of the microtome on the left-hand side and consists of
an adjustment screw for adjusting desired thickness. The cutting thickness can be read from the
cutting screw.

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Seite 11/21
CONTROLLER

Switch on the unit with the main switch at the rear.

To see and change current time and day

Push TIME key for more than 3 sec, LED switch on and HUR (hour), MIN (minutes), DAY (day)
will appear in display – to exit: push time key for more than 5 sec.

Push Set key and change the hours, minutes, day with UP / DOWN keys.

Set temperature

Push SET key and set temperature with UP / DOWN keys.

Light setting

Press LIGHT key, to switch off light press LIGHT key again.

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Manual defrost

For manual defrost press DEFROST key for more than 5 sec.

Energy saving mode

It is possible to program an energy saving mode

With the help of the energy saving modus it is possible to reduce the chamber temperature over
night at programmed hours. The on / off cycles of the compressor will be reduced. Therefore it is
necessary to set the real time correctly.

To set the energy saving times press the clock/down button for more than 3 seconds, afterwards
the ES (energy saving - night) button

ILE energy saving cycle start during workdays ( 0 - 23.5 hours ). During the energy saving
cycle the set point is increased by value of HES so that the operation is SET + HES

dLE energy saving cycle length during workdays ( 0 - 24.00 hours). sets the duration of the
energy saving cycle on workdays.

ISE energy saving cycle start on holidays ( 0 -23.5 hours )

dSE energy saving cycle length during holidays ( 0 - 24.0 hours )

HSE Temperature increase during energy saving cycle (-30 - 30°C / -54 - 54°C) sets the
increasing value of the set point during energy saving cycle.

To set the time and the weekly holidays:

press the fast freezing / up button for more than 3 sec. time LED switches on and the current
time is displayed. By pushing the SET key, the time LED will start flashing and setting the current
hour, minutes, day and the weekly holidays (up to 3) will be possible.
to exit, press SET + UP or wait 15 sec without pressing any key.

To set the energy saving times

Push the fast freezing / up button for more than 3 sec. time LED switches on and the current time
is displayed. By pushing ES key LED will start flashing and setting energy saving time will be
possible. To exit, press SET + UP or wait 15 sec without pressing any key.

To start holiday function by using the keyboard

Push the ES key for more than 6 sec. until the label Hd is shown. Time LED will start to blink
slower than during time programming and the regulators will follow holiday times.
Push the ES key for more than 6 sec. to start normal functioning again.

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PRODUCING SECTIONS

4.1 Microtome knife (cutting edge, knife angle)

The microtome knife is fixed in the knife holder of microtome with simultaneous adjustment of
relief angle. It is inserted in the suitable slot and fixed with clamp- and adjusting screws. In order
to produce good quality sections the correct fitting of knife is of great importance. The finger
protection is inserted on both sides of knife holder leaving the cutting surface free in the centre.
Then the knife is tightened by adjusting the relief angle.

Knives or disposable blades can be

used. For use of disposable blades a special blade holder is required. Better results, however,
arc obtained when using classical steel blades, in most cases C-profile. Many microtome knives
are provided with a cutting bevel to allow best cutting behaviours, since very favourable large
cutting angles are achieved.

Knife bevel of C-Knife

The features of a knife are determined by the steel quality and precise grind with even and fault
free bevel. For hard material like cartilage, bone, calcified material, special solutions hard metal
knives in D- or C-shape are used.
SLEE-microtome knives are factory-reconditioned by regular regrinding.

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4.2 Relief or clearance angle

Relief angle designates the angle between cutting bevel of knife and surface of specimen. High
quality sections require that the cutting edge move past the specimen block avoiding dose
contact of the rear of the knife with the block. Favourable results are achieved with a relief angle
between 10° and 15°. The size of relief angle largely depends on density and hardness of tissue,
the cutting temperature, the knife bevel type and the quality of knife

Schematic illustration of relief angle

4.3 Knife holder

The knife holder is for fixing and positioning of knife

Fig. 4 schematically illustrates the knife holder. The
knife is in its clamping position.

Illustration of knife holder

4.4 Adjusting and fixing of knife

By means of the fixing screw the knife is put into the desired position, the knife angle is adjusted
and the knife is fixed in the knife holder.
Place the knife carefully in the knife holder and avoid touching the edge. Then insert the finger
protection on both sides of the knife carriage teaving the cutting area free (see Fig. 5). Then use
the fixing screw until knife has reached desired angle. By optimising the relief angle with samples
the quality of cutting can be best influenced. The individual sections should have unique
appearance and repeatable thickness. There should be no deformation or destruction.

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Schematic illustration of finger protection

4.5 Anti-roll plate

The anti-roll plate counteracts the natural tendency of the section to transparent plastic to allow
viewing of the procedure.
On the right and left side there arc spacers positioning the plate in its correct position to the knife.
The anti-roll plate is lying on the knife whereby the edge aligns with the cutting edge. There
should be space between knife and anti-roll plate to allow the section to shield through and to
avoid rolling up (see Fig. 6).

Position of anti-roll plate

The knife in conjunction with the anti-roll plate adjustment has a considerable influence in cutting
quality.

4.6 Trimming

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Trimming is for advancing the specimen to be cut resp. the cutting level of the knife. By turning
the heft hand wheel (coarse advance) anti-clockwise the knife carriage moves to the specimen.
Observe even cutting movement and check if the cutting level has been reached. The return
movement is made by turning the coarse advance clockwise.

4.7 Cutting

The temperature in the cooling chamber has a decisive influence in the cutting quality. Select the
desired cutting thickness through micrometer adjustment and adjust desired temperature. Fix
specimen in specimen holder. Once desired temperature has been reached trimming can be
started. Sections of selected thickness arc made. The procedure is repeated unit the desired
cutting level is reached and quality of cutting corresponds the requirements. Clean the knife and
position the anti-roll plate by means of the knob above the heft-hand hand wheel. The cutting
procedure can start now. The section Lies fiat on the top surface of the knife. Temperatures for
some special tissues arc given in Table 1.

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OPTION: DISPOSABLE BLADE HOLDER

The MTC is equipped with a knife holder. The anti-roll plate can be used in the following
way:

As option, a disposable blade holder is available. You can easily put it into the holder as
you can see on the photo.
Additionally a small plate has to be installed for the right angle of the anti-roll plate. The
photo shows you how to mount it.

Addtional plate for blade holder

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GUIDE FOR CUTTING TEMPERATURES

Tissue type Temperature of knife [°C]

-7 to -10 -10 to –13 -13 to –16 -16 to –20 -20 to –25 -25 to –30
Bone marrow X X
Brain X
Breast, with fat X X
Breast, without fat X X
Cartilage X X
Fat X X X
Heart X
Bowel, X X
intenstines
Kidney X X
Larynx X X
Lip X X X
Liver X X
Lungs X X
Lymph node X X
Lymphatic vessel X X
Lymphoid X X
Muscle X X
Nose X X
Rectal (large X X
intensine)
Scrape X X
Skin, with fat X X
Skin , without fat X X X
Testicle X X X
Tongue X X
Spleen X
Womb X

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TROUBLE SHOOTING

Fault Possible reasons

coming off of the block Mounting of insufficient freezing

from specimen holder block not enough freezing medium used
Platform of specimen holder is dirty or greasy
Cutting surface of block is uneven
There is not enough freezing medium in the
Grooves Of the specimen holder
Cutting Sections from specimen are too thick
preparation Tissue is too cold
The advance was operated although the specimen
Was underneath the knife
Knife Relief angle not sufficient
Consecutive sections Microtome Knife and/or specimen holder not sufficiently
Show different tightened
Thickness
Irregular specimen transport, since advance spindle
Is iced up or coarse advance stift‘
Knife iced up
Knife Insufficient relief angle (tissue block touches
shoulder between cutting level and area of knife)
Adjusting angle of knife too large, tissue is scratched
Or crushed
Temperature Tissue too warm or too cold
Chamber too warm or too cold
Interval between individual sections too long
Tissue Badly mounted at specimen holder

Different thick lines in the Microtome Knife or block not correctly tightened
sections Shatter movement of microtome shaft
Knife Cutting edge not sharp enough or specimen
includes hard structures
Cutting edge too thinly ground resulting in

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Vertical stripes or lines in
Sections
Horizontal gaps in section
Crushing of sections
vibration during cutting
Tissue Badly fixed to specimen holder
Knife Cracks or other irregularities of cutting edge
Ice particles or foreign bodies sticking on
cutting edge
Guide plates top edge of plate is damaged
Ice particles or foreign bodies on top edge
Specimen Hard particles (dirt, calcium carbonate) in
specimen
Temperature Tissue block too cold and therefore brittle
Knife Adjustment angle too steep
Cutting edge dull
Surface of knife contaminated with ice, tissue
particles or foreign bodies
Guide plate Bad adjustment of guide plate
Surface of guide plate contaminated with ice,
tissue particles or foreign bodies
Spacers damaged
Temperature Tissue, knife, guide plate too warm
Cryostat temperature too high
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