Leica CryoJane User manual

CryoJane®User Manual
Customer Service 1-800-237-2772
(In continental US only)
Customer Service 1-201-343-1313
(International)

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Instrumedics grants the purchaser of the CryoJane system
the rights to use the patented CryoJane (CFSA) materials
and process.
Patent # 4,545 831

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CRYOJANE TAPE-TRANSFER
STEP-BY-STEP
INSTALLATION INSTRUCTIONS
The following items are required for installation:
Absolute Alcohol 1600 Watt Hair Dryer Gauze Gloves
01 Remove release paper
from ECU
02 Place ECU on top left
corner of cryostat
03 Using absolute alcohol and gauze
clean chamber wall
04 Heat left wall for Mech &
Oil Bath placement
05 Place Oil Bath towards the
rear of the left chamber wall
06 Route Oil Bath & Mech Cables
underneath Mech
*07 Place Mech in Chamber.
Place Mech low in chamber
near the front wall.
(Placement near the freeze
bar is ideal in most cryostats)
08 Route cables in door bar 09 Place door bar with cables
onto front chamber ledge.
If required attach Door Bar Support Strip
underneath door bar

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10 Adhere Cable holders
along cryostat top outer wall
11 ECU Rear with Cable &
Power Connectors
12 Connect Mech, Oil Bath Cables to
ECU
& Power cord to the ECU and wall outlet
Turn the ECU on
13 Mech Cable
connected to ECU
14 External-Installation
Complete
Cryo-Vac Shown
15 Internal-Installation Complete
Cryo-Vac Shown
*The standard CryoJane Mech installation is shown in Fig. 07.
The Hacker/Bright OTF and Microm HM 560 cryostats require an S-Plate
Mech and separate mounting plate for installation.
S-Plate Mech for Hacker OTF and Microm HM 560 Cryostats
Mounting Plate locks onto S-Plate Mech.
Slide Nest Entrance
The entire assembly is adhered to the left cryostat wall as shown in Fig. 07.
Note: The S-Plate Mech must have enough clearance from the front wall of the
cryostat for slides to enter the Slide Nest.

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Table of Contents
Instrumedics Frozen Section Process Overview…………… 7
Freezing Process …………………………………………………………. 7
CryoJane Process ………………………………………………………… 7
CryoJane System Components …………………………………….. 8
CryoJane ECU ……………………………………………………………. 9
CryoJane Mech Components ………………………………………. 10
CryoJane Operating Procedure …………………………………… 11
Background ………………………………………………………………. 11
Preparation ……………………………………………………………….. 11
Use ……………………………………………………………………….. 12
Steps for a Single Section ………………………………………… 12
Steps for Multiple Sections ………………………………………. 18
Summary of the Instrumedics Frozen Sectioning Process. 19
Special Fixation … … … … … … … … … … … … … … … … … … … … … … 20
Aqueous Fixation …… ………………………………………………….. 20
“Anhydrous” Fixation …………………………………………………….. 21
CryoJane Maintenance ……………………………………………….. 22
ECU ……………………………………………………………………… 22

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Mech …………………………………………………………………….. 22
Troubleshooting Guide … … … … … … … … … … … … … … … 23
Troubleshooting the Mech and ECU …………………………………….. 23
CryoJane Process Troubleshooting ………………………………………. 25
Troubleshooting Finished Slides …………………………………………. 27
Oil Bath Accessories Kit Components ………………………….. 29
Oil Bath Accessory Kit Operating Procedure ………………. 30
Background ……………………………………………………………… 30
Preparation ………………………………………………………………. 30
Use ……………………………………………………………………… 31
Storing a Tissue Block ………………………………………….. 31
Sectioning a Stored Tissue Block ………………………………. 31
Oil Bath Accessory Kit Troubleshooting Guide ………….. 32

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Instrumedics Frozen Section Process Overview
Freezing Process - Conventional Freezing vs. Snap Freezing
In the conventional freezing method, the tissue is typically frozen at about -30°C. During
this relatively slow freezing process, the ice crystals that form are usually large compared
to cellular dimensions. These crystals can cause considerable displacement and structural
damage to the tissue.
The Gentle Jane device is designed to snap-freeze the tissue in approximately 8 -10
seconds. The freezing agent can be liquid nitrogen, LN2chilled isopentane, dry ice or a
thermoelectric cooler (Peltier device). The temperature at which the tissue is frozen
should be -60°C or colder.
NOTE: Gentle Jane Snap-Freezing methods are described in a separate Snap-
Freezing User Manual.
CryoJane Process
The CryoJane process is built around three special methodologies:
1. The capture of an undistorted, thin, snap-frozen section on a special cold adhesive
tape.
2. The lamination of the captured section onto a cold glass microscope slide coated with
an ultraviolet light curable pressure sensitive adhesive.
3. The curing of the adhesive on the slide with an 8 millisecond ultraviolet flash, and the
subsequent removal of the adhesive tape, leaving a still frozen section firmly adhered
to the microscope slide.
The CryoJane system is designed to permit the facile application of these methodologies,
and can be adapted to virtually all model cryostats.

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CryoJane System Components
FIGURE 1. CryoJane System Components
1. Mechanism (Mech). See CRYOJANE MECH COMPONENTS for details.
2. Electronics Control Unit (ECU). See CRYOJANE ECU for details.
3. ½ inch diameter Mech Cable (connected to the Mech)
4. Door Channel
5. 3 Shelf Unit
6. Hand Roller
7. Adhesive Coated Slides
8. Tape Windows
9. Knife Facet Wipers
10. AC Power Cord (not shown)
OPTIONAL EQUIPMENT:
11. Protective Oil Accessories Kit
(see Oil Bath Accessories Kit sections in this manual)
12. Cryo-Vac-Away System

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(see Cryo-Vac-Away User Manual)
CryoJane ECU
Figure 2. CryoJane ECU -back
Figure 3. CryoJane ECU -front and right side
1. On/Off Switch
2. Pad LED
3. Power LED (or Bath LED if optional Oil Bath Accessory Kit is installed).
4. Flash/Pad Connector
5. Optional Oil Bath Connector
6. Fuses
7. AC Power Cord Connector
4 5
6
7
1
2
3

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CryoJane Mech Components
FIGURE 4. CryoJane Mech Components
1. Blue Pad
2. Stainless Steel Spring Clip
3. Black Knob
4. Flash Tray
5. Lid