Leica EM FC7 User manual

Leica EM FC7
Operating Manual
Version 162220032 01/2010

Important Note
Leica reserves the right to change technical specifications as well as manufacturing
processes without prior notice. Only in this way is it possible to continuously improve
the technology and manufacturing techniques used to provide our customers with
excellent products.
Any copyrights of this document are retained by Leica Mikrosysteme GmbH, Vienna.
Any reproduction of text and illustrations (or any parts thereof) by printing, photocopy-
ing, or other methods (including electronic systems and media) requires express prior
permission in writing.
Issued by:
Leica Mikrosysteme GmbH
Hernalser Hauptstrasse 219
A-1170 Vienna

Leica EM FC7
Operating Manual
Leica EM FC7 Serial Number:
Date of purchase:
For the instrument serial number, please refer to the name type label on the
back of the instrument!
Please read this instruction manual carefully
before operating the instrument.

Page 2 Operating Manual Leica EM FC7 01/10
Foreword
This Technical Documentation is intended to provide essential information about the
proper operation of the Leica EM FC7 Cryo-chamber.
This user manual describes commissioning of the Leica EM FC7 Cryo-chamber,
phased testing and adjustment of all components and movement sequences, and
restoring the basic functionality.
Service and operating staff must familiarize themselves with all components of the
system before commissioning. Particular attention must be paid to the aspect of
safety.
This user manual must be retained for future reference.
Texts, schedules and tables may not be copied, reproduced, or divulged to third
parties without our express consent.
It should be noted that this Technical Documentation does not constitute a part of
any existing, prior agreement or covenant or legal relationship.
All obligations are derived from the purchase agreement, which is also the sole
document of record regarding the terms of the warranty. Contractual provisions are
not affected by the Technical Documentation.
The documentation issued by the respective suppliers shall apply in addition to this
manufacturer's documentation.
In addition, all generally applicable legal and otherwise binding regulations for
preventing accidents and protecting the environment must be observed and
communicated.

Page 4 Operating Manual Leica EM FC7 01/10
Table of contents
1. Safety Introduction................................................................................................................................... 6
1.1 Symbols and abbreviations.................................................................................................. 10
1.2 Abbreviations ....................................................................................................................... 10
2. Identification ............................................................................................................................................ 11
2.1 Product ................................................................................................................................. 11
2.2 Manufactured by .................................................................................................................. 11
3. Introduction.............................................................................................................................................. 12
3.1 Overview .............................................................................................................................. 12
3.2 Instrument Function .............................................................................................................13
3.3 Advanced touch screen control unit..................................................................................... 14
3.4 Basic touch screen control unit ............................................................................................ 15
4. Installation and Setup............................................................................................................................ 16
4.1 Packing list ........................................................................................................................... 16
4.2 Preparation of the ultramicrotome UC7 or UC6................................................................... 18
4.3 Unpacking the cryochamber EM FC7 .................................................................................. 19
4.4 Mounting the EM FC7 onto the Ultracut UC7 ...................................................................... 22
4.5 Connecting the Cryochamber .............................................................................................. 24
4.6 Connecting the LN2hose of the pump ................................................................................. 25
4.7 Connecting the antistatic device EM CRION with discharge mode..................................... 26
4.8 Connecting the EM CRION with discharge and charge mode............................................. 26
4.9 Installing the Micromanipulator ............................................................................................ 28
4.10 Repacking to prevent damage during transportation........................................................... 29
4.11 Storage location for the instructions..................................................................................... 29
5. Operation .................................................................................................................................................. 30
5.1 Inserting the cryo specimen holder ...................................................................................... 30
5.2 Inserting knives into the knife holder.................................................................................... 30
5.3 Inserting the knife holder......................................................................................................31
5.4 Inserting the specimen ......................................................................................................... 32
5.5 Using the antistatic device EM Crion with discharge mode ................................................. 33
5.6 Using the EM Crion with discharge and charge mode......................................................... 34
5.7 Advanced touch screen control unit to operate the EM FC7 ............................................... 35
5.8 Basic touch screen control unit to operate the EM FC7 ...................................................... 39
5.9 Operating the micromanipulator........................................................................................... 41
5.10 Set of Cryotools.................................................................................................................... 46
5.10.1 Unpacking.................................................................................................................... 46
5.10.2 Device function and operation..................................................................................... 46
5.11 Refilling the Dewar ............................................................................................................... 51
5.12 Finishing work with the EM FC7 .......................................................................................... 52
5.13 Maintenance of the valves ................................................................................................... 52
6. Technical specifications....................................................................................................................... 53

Page 5 Operating Manual Leica EM FC7 01/10

Page 6 Operating Manual Leica EM FC7 01/10
1. Safety Introduction
Liquid Nitrogen (LN2)
When working with liquid nitrogen (LN2) please bear in mind LN2is extremely cold. It
boils at -196 °C. Nitrogen gas (GN2) escapes at very low temperature from the
boiling LN2. Both LN2and GN2as well as cooled elements (e.g. pipes, valves,
hoses, containers or stoppers) can cause severe frost bite and burns to the skin and
eyes.
When LN2evaporates, it expands in a ratio of 1:700. 1 litre LN2produces almost 1
m3 of GN2. Care should therefore be taken to ensure that when large quantities of
nitrogen evaporate (e.g. when transferring LN2), the room should always be well
ventilated.
Removing LN2waste: dump LN2into an outdoor pit or container filled with gravel,
where it will evaporate rapidly and safely.
GN2is odourless and tasteless and will be inhaled like air. GN2is non-toxic, but a
high GN2content in the air (> 78%) reduces the oxygen-content (< 21%) and
produces immediate fainting and deep unconsciousness without any previous
symptoms.
When there is doubt about the adequacy of ventilation, use an oxygen analyser (0
to 25% scale) to check for oxygen. The content of oxygen must not drop below
18%.
If an unconscious person stays in a low oxygen environment then death may occur.
If breathing stops, apply artificial respiration at once and notify doctor and
ambulance immediately.
For the reasons given above, never put LN2Dewars in a closed storage room or
chamber. The evaporation rate from Dewar vessels can rise to several litres a day if
they are defective due to improper handling or to natural wear over many years of
use.
Always keep the working area well ventilated.
Bring objects at room temperature carefully into contact with LN2. Initially an
insulating gas layer is formed preventing a large transfer of heat. During this initial
period little LN2evaporates. However, once the object has cooled down there may
occur unexpected strong boiling and spurting of LN2.
In the case of burns from LN2splashes, rinse the affected skin immediately with
plenty of water at hand temperature. For serious burns arrange for a skin specialist
to see them at once.
In the case of LN2affecting the eyes, rinse immediately with water at hand
temperature and arrange for an eye specialist to see it at once.
Never use glass Dewar vessels in the lab (especially glass Dewars larger than 2
litres capacity) without complete metal envelope: Glass Dewars often burst for no
obvious reason or due to unintentional mishandling (e.g. contact with metal
instruments etc.). Never work without open protective glasses when using LN2in a
glass Dewar.

Page 7 Operating Manual Leica EM FC7 01/10
Fig.1.1: When working with LN2avoid protective glasses (a), boots (c), walking
shoes (e) and protective gloves (g) out of which the LN2cannot easily escape if
entered. LN2splashing into the closed protective glasses (a), open boots (c), shoes
(e) or protective gloves (g) evaporates suddenly and can cause serious burns.
Always use protective glasses (b) with side protection which are open at the top and
at the bottom. Only use boots if you have loose (not narrow) trousers coming
outside the boots (d) and completely covering the gap. Wear only open slip-on
sandals (f) in the lab, no walking shoes or court shoes. Always wear cuffless
trousers if you wear slip-on sandals. Never wear protective gloves when pouring
LN2or when putting the Dewar head on the Dewar vessel. Just use an open flannel
cloth (h) to protect your hands from the cold. Gloves should be used only to grasp
dry cold parts. They are unsuitable for LN2work.

Page 8 Operating Manual Leica EM FC7 01/10
Only use metal Dewars specifically designated for storage of LN2, since only
containers of this kind exclude risks during storage. For routine cryopreparation
metal troughs (1 cm styrofoam insulation), styrofoam containers or plastic troughs
are eminently suitable and ensure low risk cryopreparation.
Check the evaporation rate of your metal Dewar regularly every three months and
compare these rates with the rate given by the manufacturer. The evaporation rate
of an undamaged metal Dewar should be well below 1 litre of LN2per day.
Defective Dewar vessels with higher evaporation rates are a safety risk, and should
be taken out of work or repaired.
Do not leave LN2standing in open vessels where it can exchange with the room
atmosphere. The boiling point of LN2(-196 °C) is lower than liquid oxygen’s boiling
point (-183 °C). When the exchange surfaces are extensive enough, oxygen from
the air will be taken up in exchange for nitrogen. LN2with high liquid oxygen content
has a faintly bluish colour. Concentrated liquid oxygen promotes vigorous burning!
Make sure that your Dewar vessel is filled only with LN2. Apply a note in the central
distribution place stating clearly
ONLY LIQUID NITROGEN
or similar if different liquefied gases are delivered from there. Check the colour of
cryogen: Bluish colour indicates the presence of a high percentage of liquid oxygen.
The concentration of liquid oxygen increases during long periods of storage as its
boiling point (-183 °C) is higher than the boiling point (-196 °C) of LN2.
Main supply must be assured: 100 – 240 VAC, 50 – 60 Hz
The instruments are equipped with protected ground. Before connecting it to the
local electrical supply make sure that the mains has the required ground and that
the instrument is connected to it according to the local regulations.
Unplug the instrument before installing or changing fuses.

Page 9 Operating Manual Leica EM FC7 01/10
HAZARD WARNING
LIQUID NITROGEN, LN2
Suffocation
- Any vessel containing LN2is a potential hazard
- One litre LN2produces 700 litres N2gas
- N2gas is odourless and tasteless
- Oxygen levels can quickly drop in confined spaces due to displacement of
oxygen
- by N2when using or dispensing large volumes of LN2
- This can cause immediate fainting and unconsciousness
- Always use LN2in well-ventilated areas
- Treat it with respect!
Storage
- For reasons mentioned above do not store full LN2Dewars in confined
spaces
Burns
- LN2boils at -196°C. It is extremely cold and can cause serious burns. Please
read the safety instructions provided with all Leica products for the correct
handling of liquid nitrogen!

Page 10 Operating Manual Leica EM FC7 01/10
1.1 Symbols and abbreviations
Caution!
This symbol alerts the user to important information which may endanger
staff or result in damage to the system if it is ignored.
Note!
This symbol indicates further information relating to a previous explanation,
which does not have a safety-critical function. However, it is important to
observe this information to ensure that the system functions optimally.
1.2 Abbreviations
CU = control unit
n.i. = not illustrated
E-W = East - West
LN2= Liquid Nitrogen
N-S = North - South
RC = Rapid Cooling
RT = room temperature

Page 11 Operating Manual Leica EM FC7 01/10
2. Identification
2.1 Product
Leica EM FC7 Cryo-chamber
2.2 Manufactured by
Leica Mikrosysteme GmbH
Hernalser Hauptstrasse 219
A-1170 Vienna
Tel.: [phone]
Fax: [phone]
Internet: http:// www.leica-microsystems.com

Page 12 Operating Manual Leica EM FC7 01/10
3. Introduction
3.1 Overview
The Leica EM FC7 is the low temperature sectioning system for the Leica Ultracut
UC7 (and UC6) ultramicrotome. It is used for semi- and ultrathin cryosectioning of
biological and industrial samples. This includes Frozen Hydrated Sectioning (FHS)
of vitrified specimens, cryoprotected biological samples (Tokuyasu technique) and
soft industrial specimens such as rubbers and polymers.
The Leica EM FC7 has been specifically designed for maximum ergonomics and
ease of use. It has an integrated chamber illumination, allows to use an integrated
antistatic device and provides a wide temperature range from –15 °C to – 185 °C for
cryosectioning. All controls are via the control unit (CU) of the ultramicrotome to
save workspace on the table and provide maximum comfort and ergonomics for the
operator.
The EM FC7 system consists of
· Cryochamber with integrated LED illumination
· Dewar vessel on mobile trolley
· Liquid nitrogen pump
· Power supply
· Connecting cables, LN2hose
· Accessories box containing 2 rotating knife holders, 1 standard specimen
holder, specimen carriers, special keys, tools and forceps
The „complete working outfit“contains in addition to the items mentioned above:
· Flat specimen holder
· CRION antistatic device, remote controlled
This allows controlling the ioniser from either the “Advanced touch screen
controller“or the “7” touch screen controller“of the Ultracut UC7.
Recommended accessories are
· Micromanipulator
· Filling System for LN2Dewar

Page 13 Operating Manual Leica EM FC7 01/10
Leica EM FC7 mounted on Ultracut UC7 with Advanced touch screen control unit
3.2 Instrument Function
Operation of the EM FC7 is performed via the control Unit of the Ultracut UC7.
Before quickly cooling down for trimming and sectioning, the cryochamber has to be
unpacked, mounted on the EM UC7 and connected to the Ultracut UC7 CU. As the
cryochamber is mounted on the chassis of the ultramicrotome it is isolated from
external vibrations. The contact free through-the-wall specimen arm provides
optimum temperature and mechanical stability.
After connection of the LN2hose to the LN2filled Dewar via the automatic pumping
system, the pump ensures a stable low temperature environment around the knife-
specimen area by pumping LN2into a tank in the EM FC7 wall. This LN2is then
heated and the evaporating gas is used to cool down chamber, specimen and knife.
Temperature stability of specimen, knife and gas in the chamber are individually
controlled by regulators close to knife and specimen. The desired temperature can
be set, changed and stored by the operator via the control unit while the actual
temperatures are displayed on the CU.
After finishing cryosectioning, a bake-out function allows the cryochamber to heat
up to +110 °C before returning to room temperature (RT) automatically.

Page 14 Operating Manual Leica EM FC7 01/10
3.3 Advanced touch screen control unit
After connecting and switching on the EM FC7 and the EM CRION to Advanced
touch screen control unit of the Ultramicrotome UC7, the touch screen appears as
shown below.

Page 15 Operating Manual Leica EM FC7 01/10
3.4 Basic touch screen control unit
After connecting and switching on the EM FC7 and the EM CRION to the 7” touch
sensitive control unit of the Ultracut UC7, the main screen of the controller appears
as shown below.

Page 16 Operating Manual Leica EM FC7 01/10
4. Installation and Setup
4.1 Packing list
LEICA EM FC7 Low Temperature Sectioning System contains:
EM FC7 cryochamber
Power supply, connecting cables
Pressure-less automatic filling system
Insulated, threaded connecting hose
Dewar for 25l LN2with special flange and
mobile trolley and pump storage holder
The case for accessories contains:
Specimen carriers 50 pcs. with concentric
rings
Specimen carriers 50 pcs. with slot
Allen key 3 mm
Cryotool with M4 thread.
Collet specimen clamp
Rotating knife holder 2pcs.
Preparation plate
Allen key 2mm
Special long forceps with insulation coating,
Black lever, 2 pcs
0.5mm spacer (transparent plastic foil)
Transparent cover used during the cooling
down process.
The Complete Working Outfit contains in addition:
EM CRION antistatic device (either with
discharge and charge mode or discharge
only), connecting cables

Page 17 Operating Manual Leica EM FC7 01/10
Optionally available accessories:
Micromanipulator for frozen hydrated
sectioning
Set of cryotools
Dewar Refilling System
Cryo Mesacut Mirror
Trimming Tool
Different Flat Specimen Holders

Page 18 Operating Manual Leica EM FC7 01/10
4.2 Preparation of the ultramicrotome UC7 or UC6
Remove armrest, knife support and segment
arc from the Ultracut EM UC7 or UC6.
Switch on the Ultracut.
Ultracut after removal of armrest, knife
support and segment arc.
Activate the FC Mount Function of the
Ultracut by pushing the FC Mount button of
the controller.
With the FC Mount Function of the Ultracut
UC6 or UC7, the knife stage will be centered
in the E-W direction and to the outmost south
direction automatically. The backlight will
switch off, the toplight will switch on. This will
facilitate mounting the EM FC7.

Page 19 Operating Manual Leica EM FC7 01/10
4.3 Unpacking the cryochamber EM FC7
Open box and lift out EM FC7 cryochamber and LN2hose: