Leica CryoJane User Manual

CryoJane® User Manual

Customer Service 1-800-237-2772

(In continental US only)

Customer Service 1-201-343-1313

(International)

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

04 Heat left wall for Mech &

Oil Bath placement

*07 Place Mech in Chamber.

Place Mech low in chamber

near the front wall.

(Placement near the freeze

bar is ideal in most cryostats)

02 Place ECU on top left

corner of cryostat

05 Place Oil Bath towards the

rear of the left chamber wall

08 Route cables in door bar

03 Using absolute alcohol and gauze

clean chamber wall

06 Route Oil Bath & Mech Cables

underneath Mech

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

15 Internal-Installation Complete

Cryo-Vac Shown

13 Mech Cable

connected to ECU

14 External-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 Troubl eshooting ………………………………………. 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, LN2 chilled 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 Snap­Freezing 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)

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7

1 2 3

CryoJane ECU

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Figure 2. CryoJane ECU - back

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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

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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

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CryoJane Operating Procedure

Background

In conventional sectioning the operator usually guides the cut section over the knife using
a brush or an anti-roll device. Even with a skillful, steady hand, this step can be difficult,
especially if a 2 to 4 micron thick section is desired.

To mount the section, a room-temperature slide is touched to it. When contact is made
between the warm slide and the frozen section, instantaneous melting of the section
occurs, converting the ice crystals back into water. Surface tension causes the melted
section to adhere to the slide. When the ice melts, water fills the spaces formerly
occupied by the ice crystals, rehydrating the nucleoplasm and cytoplasm, causing
erythrocytes to lyse and soluble substances to be displaced. Flow of the melted solutions
can distort and displace fine structure. Often the melted section is then dried, during
which step, surface tension forces further distort, displace and collapse tissue structure.
Pathologists and clinicians have been trained to recognize normal and abnormal structure
against this background of degraded fine structure.

In contrast, the CryoJane process is designed specifically to avoid melting and air drying
and its associated artifacts. It uses a cold adhesive tape to support and capture the section
during the cutting step. The unthawed section on the tape is then transferred to a special
cold adhesive-coated slide. The section is not melted in the mounting step. The
adhesive coating on the slide is then polymerized, and the tape is peeled away, leaving the
section permanently anchored to the slide while still frozen.

Preparation

• A day’s supply of Adhesive Coated Slides and Tape Windows should be in the

cryostat, chilled and ready for use.

• The cryostat temperature should be -25°C to -30°C (depending on the cryostat

model).

• The Pad light (left LED) on the ECU should be green.

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