PRECISIONARY Compresstome VF-500-0Z Vibrating Microtome User Manual

Compresstome® VF-500-0Z
Vibrating Microtome
User Manual Compresstome® VF-500-0Z User Manual

INTRODUCTION

Congratulations on your purchase of a Compresstome® VF-500-0Z from Precisionary Instruments! The VF-500-0Z tissue slicer model is the newly developed product in the Compresstome® slicer family with patented Auto Zero-Z® technology. The VF-500-0Z is fully automated to section both fixed and live tissues with a thickness range of 2 µm to 2000 µm with an adjustable precision of 1 µm. The Compresstome® VF-500-0Z excels at sectioning slices used for:

• Electrophysiology
• Immunohistochemistry
• Organotypic culture slices
• Precision cut tissue slices
• Sectioning of mature or older brain tissues
• And much more!

At Precisionary Instruments, we are thrilled to help you get started with the new vibrating microtome, and we take great pride in quality customer service. Please read the following manual to help you get started with the Compresstome® VF-500-0Z.

CONTACT US

We have multiple ways for you to contact us:
E-mail: info@precisionary.com
Phone:  Customer Service & Quotes: 617-682-0586
Technical Support: 508-810-0111
Fax: 1-866-424-2217
Mailing address:
Precisionary Instruments
207 Union Street
Ste 2
Natick, MA 01760
Training videos
To supplement this user manual, full visual training videos for all steps in successfully using the Compresstome® slicer are available online.
Our company’s YouTube account site is called, “Precisionary Instruments LLC.” You can find all of these videos in our account. The videos can also be found on our website
(www.precisionary.com) and include instructions for:
Setting up the Compresstome®
https://www.youtube.com/watch?v=q-fhdkCseyc&t=15s
Using the motor box on the Compresstome ®
https://www.youtube.com/watch?v=IbbMAdWM6p8
Using the control box on for the Compresstome®
https://www.youtube.com/watch?v=LlNkgZSwMOk
Adjusting the blade holder angle & height on the Compresstome®
https://youtu.be/p2l-0rmrYmM
Specimen tube insertion on the Compresstome®
https://www.youtube.com/watch?v=SYOpr9mX-H4
Making the agarose solution for tissue embedding
https://www.youtube.com/watch?v=0CW5j5AG4w8
Embedding tissue samples into specimen tubes
https://youtu.be/lDmodsRS8s0
Preparing tissue transfer pipettes for moving tissue slices
https://www.youtube.com/watch?v=4Sh-0TwJbKU
Transferring tissue slices cut with Compresstome®
https://www.youtube.com/watch?v=dbHuRX9eigA
Compresstome® machine cleaning
https://www.youtube.com/watch?v=x7Pe2D_uJkM
Machine oil maintenance for the Compresstome®
https://www.youtube.com/watch?v=QEDwy2eK9yg
Blade removal and clean up for the Compresstome®
https://www.youtube.com/watch?v=IXqXNHCMlQ0

ANATOMY OF THE COMPRESSTOME® VF-500-0Z

LIST OF VF-500-0Z COMPONENTS

All Compresstome® parts can be ordered online: http://precisionary.com/e-store/
Contact us at info@precisionary.com and we will send you a quote.

sectioning.
4| Vibration head| VF-VHU-VM-500-0Z| Blade oscillation assembly, which includes a blade holder, and an oscillation motor.
5| Wire to controller box| —| Connects the motor box to the controller box.
6| Specimen tube| VF-SPS-VM-12.5-NC
VF-SPS-VM-15.5-NC| Holds the specimen for sectioning and embeds tissue in agarose.
7| Motor box| —| Automated motor box that pushes and drives the specimen tube for sectioning slice to slice.
8| FF/FR controller| —| Fast Forward and Fast Reverse dial for more rapid control of the specimen tube.
9| Motor Box Plunger| —| Aligns with tube for advancement

LIST OF STARTER KIT COMPONENTS

All of our Compresstome® consumables can be ordered online at:http://precisionary.com/e-store/
Contact us directly at info@precisionary.com, and we will sent you a quote.

This is low melting agarose for embedding the tissue.
Blades| VF-BL-VM-SSB| Included are 10 double-edged stainless steel blades.
Pipettes| __

| Plastic disposable pipettes are included for easy transfer of agarose into the specimen tubes for embedding.
Glue| VF-VM-GLUE| Used for securing tissue specimens to the specimen tube.
Forceps|

__

| Small forceps for easy manual manipulation of tissue sample.
Machine Oil| VF-VM-OIL| Clear, colorless, and environmentally friendly machine oil for maintenance.
Recommended use: 1X/week apply to all moving and oscillating parts of tissue slicer, then run machine for 5-10 minutes continuously.

SETTING UP THE COMPRESSTOME® SLICER

Please remove the transportation fixture before your first-time use. Be sure to save the transportation fixture, because the fixture will need to be mounted back to the machine before any future shipment.
* IMPORTANT NOTE:
Before using the Compresstome® slicer, you MUST remove the “transportation stabilization plate” on the machine. The small metal plate is screwed in at the back of the vibrating head and is there to help stabilize the Compresstome® during shipment. Failure to remove the transportation stabilization plate may result in damage to the vibrating head.
The transportation fixture MUST be unscrewed and removed before use!

OPERATION OF THE COMPRESSTOME®

MOUNTING THE CUTTING BLADE TO THE BLADE HOLDER

For each double-edged stainless steel blade, cut the blade horizontally to form two separate blades. You will only need one of the two resulting blades for each use. For ceramic blades, remove one of the blades carefully from the pack.
Squeeze a small amount of super glue onto a petri dish or other solid surface. Pipette 5 µL of the super glue onto the blade holder.

Position the cutting blade onto the blade holder. For the double edge blade, you may notice that along the edge where you cut them, the edge may be curled up. Cut the  urled edges of the blade before gluing.
CAUTION! Make sure that you do not touch the blade edge! Allow the glue to dry for 3 minutes before use. Make sure there is no glue on blade to avoid tissue sticking to the blade.

After the glue dries, you are ready to use the blade holder and blade for sectioning. Remember: The blade holder needs to becleaned with acetone between each blade change, so that the glue residue does not build up.

MOUNTING THE CUTTING BLADE TO THE MAGNETIC BLADE HOLDER

The magnetic blade holder is used without glue for cutting with stainless- steel blades only. The magnetic blade holder can still hold ceramic and tungsten carbide blades which will need to be applied via glue. To clean the magnetic blade holder, soak in liquid overnight and dry & clean right after use.

The new blade holder will have visible magnets along the outer side (left photo) of the holder.  Locate the inner side of the blade holder (right photo), with the raised lip along the top edge, as this is the side ofthe blade holder the blade will be applied to.|
Carefully remove and cut one of the double-edged stainless-steel blades foryour experiment, and prepare to position the blade on top of the flat side of the blade holder.|
Carefully place the blade on the blade holder so that the top edge of the blade touches the bottom edge of the raised blade holder lip.|
Carefully slide the blade towards the center of the blade holder to center theblade along  he magnets.  Once your blade is centered and aligned, you are ready to use your  ewmagnetic blade holder!|

CHANGING THE CUTTING BLADE

The sharpness of the blade can directly affect the quality of the slices. We recommend that double edge razor blades be changed after cutting 3-5 samples (examples: brain, lung, gastrointestinal, and skin tissues). If tissues are more fibrous, blades should be changed after cutting 1-2 samples (examples: lymph nodes, liver, spleen, and kidney tissues).
Ceramic blade typically lasts 2-3 months depending on the degree of usage. Double edge razor blades and ceramic blades are ideal for this microtome, and these blades are what we include in the Compresstome® Starter Kit. The blades should be cleaned with acetone and ethanol before use.
Follow these instructions on changing blades for the VF-500-0Z.
To remove the blade holder from the vibrating head unit, loosen the small screw on top of the blade holder with a small Allen wrench, and remove blade holder from the axial bar.

Remove the old cutting blade and dispose of it safely intoa sharps container
If you are using the magnetic blade holder, you may carefully remove this blade by sliding it to either side of the blade holder edge, and then carefully remove the blade from the magnet.

Soak the blade holder in the acetone bath for 5 minutes. Wipe off any residual glue on the blade holder using a paper towel.
DISCLAIMER – Please only use acetone to clean the blade holder and specimen tube. The buffer tray is not made of Acetone safe materials.

Follow the instructions under “Mounting the Cutting Blade onto the Blade Holder” to install a new cutting blade.
See page 9.

PREPARING THE AGAROSE FOR EMBEDDING:

We recommend using agarose type-Ib, or low melting point agarose. All tissue specimens should be embedded in agarose. Type-Ib agarose powder can also be purchased through Sigma Aldrich at: http://www.sigmaaldrich.com/catalog/product/sial/a0576?lang=en&region=US
The following procedure assumes you are using a 0.5 mg agarose tablet. This table summarizes the buffer volumes to use for agarose tablets. We recommend using a 2.0% or
2.5% agarose solution.

Agarose Tablet Chart

Solution volume needed to achieve % agarose (mL)
Agarose %| 1 tablet| 2 tablets| 3 tablets
2.%| 33| 67| 100
2.%| 28| 56| 83
2.%| 25| 50| 75
2.%| 23| 45| 68
3.%| 20| 40| 60
3.%| 17| 33| 50
3.%| 16| 31| 47
4.%| 14| 29| 43

Each tablet contains is 0.5 mg of agarose. Depending on the gel strength you want for slicing, you can add the appropriate amount of water or buffer solution to the glass vial. We recommend adding 20-25 mL of solution to each glass vial containing 1 agarose tablet.

Dissolve the agarose tablet for 2 minutes by swirling the glass vial.

Heat the agarose as you normally would, which can be done in a microwave. Microwave the agarose solution for 10 seconds, then shake to mix it up, and repeat until the agarose solution has become clear. Heating the solution in 10 seconds bouts helps prevent the solution from overflowing. You may see a lot of bubbles in the solution, which is normal.

1. We recommend using agarose that has a low transition temperature, which means that the agarose will remain in a “liquid-like” state without congealing at low  emperatures. This way, keeping melted agarose in a warm water bath will allow you to have readymade agarose when sectioning several different specimens.
2. dvantages of using our Compresstome® agarose preparation:
   a) Eliminates weighing agarose for individual experiments
   b) Agarose tablets are fast dissolving in just 2 minutes
   c) Environmentally friendly with no organic solvents that could harm tissues
   d) Produces consistent, reproducible gels that are clear

EMBEDDING TISSUE SAMPLES IN AGAROSE:

Place the syringe chilling block into water and icebath or a freezer to cool (usually at least 15 min).| Try cooling the chilling block at -20 °C (put it in the freezer for 10 min)
---|---
Prepare your tissue sample by cutting it so that the tissue fits inside the specimen tube.|
Squeeze a small amount of super glue onto the specimen tube base.|
Using forceps, position the tissue onto the specimen tube base and glue your tissue to the base.| ![PRECISIONARY Compresstome VF 500 0Z Vibrating Microtome

• ANATOMY OF THE COMPRESSTOME®16](https://manuals.plus/wp- content/uploads/2022/11/PRECISIONARY-Compresstome-VF-500-0Z-Vibrating- Microtome-ANATOMY-OF-THE-COMPRESSTOME®16.png)
  Place the embedding cap onto the top of the specimen tube.|
  Pipette enough agarose to fully cover the tissue sample. Gently tap on the syringe cap to dispel any air bubbles.|
  Withdraw the syringe tube downwards so that the tissue sample enters the tube. You can then
  remove the syringe cap.|
  Place the syringe chilling block (should be prechilled, see step  1)over the specimen tube to chill
  the entire sample and help the agarose  solidify. This process usually takes less than a minute,
  and  at most 1.5 minutes.|
  Once your tissue is embedded into agarose, you are ready for sectioning with the Compresstome® slicer!|

SECTIONING WITH THE COMPRESSTOME® SLICER:

Connect the supplied power cable to the control box and the control box to the VF500-0Z  microtome. Makesure the connections are good by tightening both screws on the connector or the machine willnot work properly. Turn the power on. 

Insert the specimen syringe tube into the buffer tray on the Compresstome® slicer.

Push the tube as far as it will go into the buffer tray with the tapered end going first. A small stopper knob on the tube will prevent it from sliding all the way into the buffer tray. (Note that your blade should have already been aligned at this point to save time during the slicing process).

Continue cutting with the Compresstome® slicer. You can adjust the slice thickness, sectioning speed, and frequency of oscillation to best suite your experimental needs.

CLEANING UP THE COMPRESSTOME® VF-500-0Z:

Empty buffer from the buffer tank.
DISCLAIMER – Please only use acetone to clean the blade holder and specimen tube. The buffer tray is not made of Acetone safe materials. We ecommend using 70% ethanol solution or 10% bleach solution to clean the buffer tray.

Loosen thumbscrew and push the step motor drive away.

Remove the specimen syringe from the plastic base.

Remove plunger from the metal syringe and scrape residual glue from the front end of the plunger.

Clean up any debris on the plunger and inside of the syringe tube with acetone swabs or soft brush.
DISCLAIMER – Please only use acetone to clean the blade holder and specimen tube. The buffer tray is not made of Acetone safe materials.

Remove razor blade from blade holder.

Dip the blade holder into the acetone bath for 5 minutes. Clean up the residual glue on the blade holder with acetone swabs.
DISCLAIMER – Please only use acetone to clean the blade holder and specimen tube. The buffer tray is not made of Acetone safe materials.

THE AUTO ZERO-Z® TECHNOLOGY

Auto Zero-Z® is our latest breakthrough technology at Precisionary Instruments. This is a patented feature designed to allow the blade holder to operate in near zero Z-axis deflection without the need to optically align every single time.
Before arriving to your lab, your Compresstome® slicer is precisely aligned in the factory to work in the optimal condition (zero ΔZ). Please ensure that you follow the special instructions for blade mounting to ensure that Auto Zero-Z® works every time you slice.
The advantages of the Auto Zero-Z® combined with Compresstome® techniques are:

• Healthier slices and better surface structure preservation.
• Thinner minimum slice thickness. By combining the Auto Zero-Z® and Compresstome® technology it is now possible to achieve an unprecedented fixed brain slice thickness of 2 µm without paraffin embedding or freezing.
• No vibration marks on the fixed brain slices. The result is a very smooth and flat slice surface which is optimal for histological processing.
• No optical alignment device is required for Zero-Z operation.
• No blade alignment procedure is required when changing to a new blade.

WARRANTY
There is a one (1) year warranty for the VF-500-0Z microtome. Additional years of warranty are available for purchase. All demos for Compresstome® slicers and consumables are considered to last 30 days from date of delivery (arrival). Consumables and accessories are non-returnable after any use. Delivery is considered to be completed when items arrive to the customer. Acceptance of items is deemed complete after 7 days of delivery (arrival), with no further signature required. Tampering with the VF-500-0Z model (the control box, step motor box/micrometer, or vibrating head unit) in an attempt to repair without written consent from Precisionary Instruments within the first year of purchase voids the warranty.
PACKING THE COMPRESSTOME
When you need to ship the VF-500-0Z back to us for maintenance or repair, please keep these three points in mind:
a) Put the transportation fixture back to the machine and tighten the screws. This will prevent damage during shipping. Failure to do so may cause damage to the vibrating head unit.
b) The FF/FB switch on the stepper motor box is a delicate part. Do not put heavy things, such as control unit, chilling block, on top of it. That will damage the switch during shipping.
c) Wrap the control unit, chilling block and power adaptor with foam sheets or bubble sheets and put them into small cardboard boxes. These will prevent paint damages during shipping.

CONTACT INFORMATION

Additional questions? Want some assistance?
We have multiple ways for you to contact us, including
E-mail: info@precisionary.com
Phone:
Customer Service & Quotes: 617-682-0586
Technical Support: 508-810-0111
Fax: 1-866-424-2217
Mailing address:
Precisionary Instruments
207 Union Street Ste 2
Natick, MA 01760 20
Precisionary Instruments © 2022

References

• Compresstome & Microtome Store | Precisionary Instruments
• Vibrating & Rotary Microtomes | Precisionary Instruments
• Agarose Low EEO 9012-36-6

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