Answers to your questions about Compresstomes, rotary microtomes, and getting better tissue slices
Your sample sits on the plastic plunger and gets pulled down within the metal tube to then be embedded with a low-melting agarose. The agarose is then chilled with a chilling block and the agarose assists with the compression technology. As the agarose-embedded sample exits the tube, the blade comes down and makes a cut right at that site of compression. This is how the Compresstome® creates even, consistent slices every time you cut.
The best cutting parameters to use for cutting speed and oscillation frequency are:
These parameters work best for cutting fixed tissue, and will not yield any chattermarks! If these parameters do not suit your needs, contact us today! We will be happy to discuss with you what works best based on your experiments and what you are cutting.
The Compresstome is a unique tissue slicer with patented compression technology. It was made to produce slices faster than other market slicers without chattermarks. If you are getting chattermarks on your tissue slices, there are a few diagnostic keys you can try:
1. Set the speed lower (try “Advance” setting of 1-1.5) and set the oscillation frequency higher (try setting of 3-4).
2. Make sure that you are using 2-3% agarose, and that there are no air bubbles in the agarose surrounding the tissue.
3. Ensure that the blade is lined up correctly with the specimen tube (within a few microns from it). If the blade is too far from the specimen tube, this will disrupt your slices.
4. Try changing the cutting blade in case it has nicks on the edge that interfere with cutting.
5. If you are working with tissue that have open cavities inside, make sure to inject/infuse the cavities with agarose or gelatin to ensure that all air spaces have been filled up.
If you cannot diagnose the problem based on these tips, give us a call or email and we will be happy to help!
If you are getting uneven tissue slices of inconsistent thickness, it may be that the ratio of agarose density to tissue density does not match. For example, agarose that is too soft (<2%) will not provide enough stability for firmer tissue (i.e. cardiac or lymph node tissue which is more fibrous) when being cut. This in result could produce chatter marks in slices. If this is happening, use a higher percent of agarose to accommodate the tissue (3%). Another fix for inconsistent slice thickness will be to lower the cutting speed (“Advance” to 1-1.5) and to increase the oscillation frequency (try 4-5).
The advantages of the Compresstome outweigh other market slicers for many reasons. These reasons include:
The Compresstome tissue slicer has many advantages over other tissue slicers. With a wide range of slicing capabilities, it is ideal for many tissue types, animal types, and experiments. Contact us today to see how the Compresstome tissue slicer can benefit you!
Our VF-510-0Z is our most popular Compresstome® model. The VF-510-0Z is our newest Compresstome model and is fully automated. Our vibratome comes calibrated with Auto Zero-Z patented technology, which gives you healthier live slices and smoother fixed sections**. This model comes with a one-touch unlocking mechanism on the sliding control box that allows for rapid sample loading and cutting, saving you time.
**Our Compresstome VF-510-0Z vibratome is equivalent to the Leica VT1200S model. Our Auto Zero-Z Technology is patented and is equivalent to the VibroCheck. However, our Auto Zero-Z Technology is already included in the price of your VF-510-0Z!
The biggest differences among these models are the automation, warranty time, and Auto Zero-Z technology.
Compresstomes® with Auto Zero-Z technology have Z-axis vibration of less than 1 micron, as confirmed by a precision laser measuring system with 0.25 micron resolution. This low Z-axis vibration is possible due to the use of precision manufactured components, 100% inspection, and careful assembly methods. Unlike other slicers there are no adjustments to slip, and no Z-axis calibration necessary by the user. The Compresstome models VF-210-0Z, VF-510-0Z, and VF-800-0Z have this Auto Zero-Z technology.
The Compresstome® VF-300 and VF-510-0Z models are fully automated models, which means that there are both ‘Single’ and ‘Continuous’ cutting modes when operating them. The micrometer sits in the control box and automatically advances your sample forward to be cut, depending on the slice thickness display that the user sets on the control box. These units can cut either one slice at a time or continuous slices and the mode is easily set via a switch on the control box. For these reasons, the VF-300 and VF-510-0Z models are excellent options for producing super thin sections, down to ~10 µm however the VF-300 only has a 1-year warranty whereas the VF-510-0Z has an industry leading 5-year warranty.
The VF-510-0Z has a sliding control box for easier access to removing and inserting the specimen tube. This means the buffer tray and the blade holder stays in place while you load the specimen. Additionally, the VF-510-0Z also has an oscillating motor cover to protect those oscillating parts from any aerosolized buffer solution coming in contact with it. This assists in the longevity of those moving parts. Because the control box is smaller in the VF-510-0Z, there is no display for the slice count or the total µm distance.
On the other hand, the VF-210-0Z is a semi-automated model meaning it can only cut one slice at a time. There is no continuous cutting mode as this model’s micrometer is a manual dial that the user needs to turn to their desired slice thickness in between each slice that is produced. Because of this manual micrometer, this model is recommended for fresh tissue slicing or producing sections >20 µm. The VF-210-0Z also has a 5-year warranty.
The VF-800-0Z is a vibrating microtome that is designed for sectioning the tissue of larger samples. It is also the only vibratome that can section large organs without freezing. With a diameter of up to 100mm, it allows the brain sectioning of cats, dogs, pigs, and monkeys. Many neuroanatomy and neuropathology experiments require sectioning entire brains from these animals. The VF-800-0Z is capable of sectioning slices for these large primates down to 40 microns. It produces even and consistent slices and it’s 5X faster than other vibratomes on the market. It also has our patented compression technology and agarose embedding process that eliminates chattermarks. The patented Auto Zero-Z technology reduces Z-axis deflection to <4 μm. The VF-800-0Z has a 1-year warranty.
Both of these models are fully automated, utilize a sliding control box for ease of loading and unloading the specimen tube and are built under strict QA procedures. The most significant differences between these models is the Auto Zero-Z technology and the warranty time, and their recommended experimental applications.
The VF-510-0Z is precisely aligned during manufacturing to eliminate any deviations in the Z-axis during the cutting process. This patented technology results in “zero Z-axis” stability that helps to produce slices with smooth surfaces that are even and filled with healthy cells. Another key feature of the VF-510-0Z is the oscillating motor cover which protects the oscillating parts from any aerosolized buffer solution coming in contact with it. This assists in the longevity of those moving parts.
In addition, the VF-510-0Z has a 5-year warranty which is the best in the industry. The VF-300 has a 1 year warranty which is an industry standard.
Finally, we recommend the VF-510-0Z as the key model for electrophysiology experiments, precision-cut tissue slices, and organotypic culture slices.
The VF-210-0Z, VF-300, and VF-510-0Z models all accommodate either a 15.5 mm or 20 mm diameter specimen tube. Each Compresstome® order includes two (2) specimen tubes of the same size. We also sell additional specimen tubes and specimen tube kits for purchase, which include two specimen tubes, one buffer tray, and one chilling block of the same-size diameter.
For larger tissue samples, such as macaque brain, our VF-800-0Z model is made with either a 75 mm or 100 mm diameter specimen tube and this size is fixed on the machine.
For the Compresstome® VF-210-0Z, VF-300, and VF-510-0Z models, the maximum length of tissue you can fit within the specimen tube is 2.5 cm.
If the white plunger gets stuck inside the stainless steel specimen tube, do not worry! This is an easy fix, all you need is a few drops of acetone. Simply drop a few drops of acetone into the specimen tube, or take a paper towel dipped in acetone and scrub the specimen tube. This will clean the glue off the plunger, releasing it from the specimen tube!
The white plunger gets stuck when an excess of glue builds up on the plunger. To avoid this getting stuck in the future, make sure you are only using a drop or two of glue when gluing your tissues onto the white plunger. We recommend 5 µL of glue to keep the tissue on the white plunger, without it building up over time.
Yes, agarose embedding is safe when using the Compresstome®! This has been our protocol for over 15 years and has been tried and true in over hundreds of labs all around the globe. We provide low-melting agarose tablets with all Compresstome® orders (in our Starter Kit of consumables) as well as for purchase in quantities of 10, 20, 50 and 100 tablets. The agarose embedding is safe for all tissue types because of our protocol to use the chilling block immediately after embedding your sample within the metal tube. The chilling block gels the agarose and takes ~30 seconds or less to do; while keeping your sample at biologically safe temperatures throughout the entire process.
We do not recommend using the Compresstome® without agarose embedding as the agarose provides 360˚ stability within the specimen tube, aids in the compression technology, and ultimately assists in producing the high-quality sections the Compresstome® cuts.
The temperature of the agarose will not damage your tissue. After heating up your agarose solution in 10 second bouts (heat, then shake, heat again & shake if necessary until the agarose is dissolved), it will need to be cooled for 5 minutes in a 32 degree C bath. Using agarose with a low transition temperature will keep the agarose in a “liquid-like” state, without it congealing at low temperatures. This is ideal for preparing your tissue samples, because the agarose will be ready when it’s time to section the possible several different specimens for sectioning. The agarose tablets we provide are ideal for embedding tissue samples, as they help produce consistent slices throughout sectioning and eliminate weighing agarose for individual experiments.
The agarose concentration needed for embedding tissue depends on the volume of buffer or water solution you want to use. If you want a stronger gel strength, you will add a higher concentration of agarose.
We provide 10 agarose tablets with our Starter Kits (also can be found in our E-Store). Each tablet contains 0.5 mg of agarose. We recommend 1 tablet of agarose for 25 mL of buffer/water solution, which gives you 2.0% concentration of agarose in your solution for embedding.
Using less or more buffer/water solution? Not a problem! We are happy to assist you in finding the concentration of agarose that suits your embedding needs. Contact us today!
There are a couple of tricks that you can try to get your tissue slices to fall out of the agarose after they are cut. The first is to use a higher concentration of agarose, such as 3% or 3.5%. If the agarose concentration is firmer, then it is more likely to automatically fall off the tissue after each slice is cut. The second option is to brush your tissue with a sucrose solution before embedding it in the agarose. Coating the tissue with sucrose gives the tissue an extra layer that allows the agarose to fall off easier after it is cut.
Yes, you can do that! Unlike the procedure of detaching the agarose from the tissue, keeping the agarose rim around the tissue is quite simple. To preserve the agarose rim around each tissue slice, use a slightly lower concentration of agarose, 1.5% to 1.8%. Keeping the agarose rim around the tissue helps keep the slices protected after cutting. This also allows for easy maneuvering of tissue slices to mount on slides without damaging the tissue itself.
There are several ways to clean the Compresstome® and the parts of the machine. Including:
Yes! Our stainless steel buffer tray is available on our E-Store for purchase and is autoclavable. You may also autoclave the blade holders.
When cutting with the Compresstome®, the sections fall into a buffer tray as they are free floating. There are several ways to keep the solution cold during cutting, including:
You want the solution in the buffer tray to fully cover the specimen tube in order to produce the best results, which means that the buffer tray will be filled to the brim with solution. You will need 210 mL of solution to fill the buffer tray when working with the 20 mm specimen tubes, and 215 mL of solution to fill the buffer tray when working with the 15.5 mm specimen tubes.
The solution you choose to use is based on your experiments. While PBS is the most commonly used buffer solution, many who are cutting fresh brain tissue will use aCSF, or those slicing liver or kidney will use KHB solution.
The simple answer is, contact us! We are happy to help troubleshoot your cutting parameters, as many low-quality sectioning issues are due to:
If you find that the issues persist after adjusting the cutting parameters, please call or email us with a description of the problem and the serial number of the model ready. Repairs typically take 1-2 weeks to complete and send back to the customer upon receipt of the model in our facilities.
For large pieces of tissue, we recommend the VF-800-0Z. It is our largest Compresstome model. Our VF-800-0Z is the only vibrating microtome in the industry that can slice larger tissue, such as sectioning of non-human primate brains, without the need to freeze the tissue. The diameter of the VF-800-0Z is nearly 100 mm and has a depth of 90 mm, which can help with neuroanatomy and neuropathology research for whole slicing from up to 2000 µm down to 40 µm. The tissue is sliced horizontally with a vibrating razor blade at a 20 degree angle at a single or continuous cutting mode. We offer a 1-year warranty for the VF-800-0Z and our patented Auto Zero-Z technology is included. This means that there is no Z-axis calibration required by the user which helps speed up the slicing process.
The VF-800-0Z is a vibrating microtome that is designed for sectioning the tissue of larger samples. It is also the only vibratome that can section large without freezing. With a diameter of up to 100 mm, it allows the fresh brain tissue sectioning of cats, dogs, pigs, and monkeys. Many neuroanatomy and neuropathology experiments require sectioning entire brains from these animals. The VF-800-0Z is capable of sectioning slices for these large primates down to 40 microns. It produces even and consistent slices and it’s 5X faster than other vibratomes on the market meaning that it has the ability to produce large through-put sections. It also has our patented compression technology and agarose embedding process that eliminates chattermarks and reduces Z-axis deflection to <4 μm.
There are two types of blades we offer for cutting tissue. We recommend ceramic blades and stainless steel blades. Both types of blades are ideal for cutting tissue. You can purchase both of these blades from our E-Store.
Ceramic blades are high quality, durable and long-lasting. They usually only need to be replaced once a month, more or less depending on how often they are being used. They are sold as a single blade unit.
https://precisionary.com/product/compresstome-cutting-ceramic-blades/
The stainless steel blades we recommend are double-edged blades for cutting tissue. The double-edge allows for double use. They are high quality, durable blades. These blades should be replaced after every use for best cutting results. They are sold in a 10 pack of blades.
https://precisionary.com/product/compresstome-cutting-stainless-steel-blades/
Stainless steel blades and ceramic blades work just the same for your slicing needs. The difference is how often these blades need to be replaced. Stainless steel blades need to be swapped out after every experiment to yield best results. They can last up to 3-5 days at most depending on the frequency of use of your Compresstome. On the other hand, ceramic blades can last up to 3 weeks depending on Compresstome use. We highly recommend these blades, as they last much, much longer than stainless steel blades. The disadvantage of ceramic blades is that if any slight nicking appears on the blade surface, this can dramatically impact your slices.
Rotary microtomes are routinely used for making paraffin-wax embedded tissue sections for histology and pathology. Because they are used for high volume sectioning, rotary microtomes need to be robust.
Precisionary Instruments offers a full line of high quality rotary microtomes for histopathology, research, and clinical purposes. Our microtomes give you reliable sections over a range of tissue slice thicknesses.
A rotary microtome is a type of microtome that cuts thin sections by moving the sample up and down across a cutting knife. The samples are usually held in a specimen head. The up-and-down movement of the specimen head is controlled by a handwheel—hence that “rotary” name! Rotary microtomes are typically used for cutting tissue samples embedded in paraffin wax. These are usually tissues obtained from surgical biopsies for histopathological analysis. Thus, rotary microtomes are commonly used in histology and pathology studies.
To operate a rotary microtome, you would first insert a low-profile or high-profile blade into the blade holder on the knife block. Then, make sure the blade guard is lifted to protect your hands from the blade. Next, insert your paraffin wax cartridge containing your sample into the specimen clamp. Then, you will use either a coarse wheel (in manual models) or press a button (in semi-automatic and automatic models) to advance the specimen clamp toward your knife block. Select the desired thickness of your slices and start sectioning to your desired area.
Yes, here at Precisionary Instruments, we currently carry three (3) types of rotary microtomes. The RF-600 which is our manual model, the RF-800 which is our semi-automatic model, and the RF-1000 which is our fully automated model. More information on those models can be found below.
We carry three (3) types of rotary microtomes. These are the RF-600, RF-800, and RF-1000 models.
There are additional differences in the technical specifications of these 3 rotary microtome models, which you can explore in detail here.
We offer a full three (3) year warranty that covers all rotary microtome models. The warranty covers parts, shipping, repairs, and labor fees should there be any defect in the rotary microtome unit that is due to manufacturing. However, the warranty does not cover any defects resulting from improper use.
Yes! We offer an annual service plan that covers your rotary microtome’s maintenance and care after your warranty period ends. The Annual Service Plan comes in two different tiers: 1) Basic Plan, and 2) Premium Plan. Each annual service plan covers all repair labor fees and maintenance costs, including parts, accessories, and shipping fees. Think of the annual plan as providing an extended year of warranty coverage. This helps give you peace of mind! You can read more about the Annual Service Plan Agreements here.
There are three different consumables needed to operate your rotary microtome
A hot water bath is essentially a paraffin section flotation bath that helps handle sectioned samples in histology and pathology laboratories. The hot water bath is filled with hot or warm distilled water, which allows for meticulous manipulation of cut tissue sections onto glass slides. These flotation baths are designed to prevent wrinkling and distortion of thin paraffin-wax embedded sections after they are cut and before they are mounted onto glass slides.
The knife block comes with 3 components. These components are below:
Our microtomes are designed for easy cleaning. The waste tray can be easily cleaned out by sliding it off of the base of the microtome and disposing of the waste into a biohazard waste bin. We recommend cleaning the tray and machine with 70% isopropyl alcohol and to allow the machine to fully dry before the next use.
ParaGuard Paraffin Repellent is recommended to remove paraffin residue to prevent buildup.
In the unlikely event that your rotary microtome needs assistance, reach out to us at Precisionary Instruments first. You can do so through our contact form, or through email, telephone, and WhatsApp. A real person will respond to you and we will directly assist you.
We respond to inquiries within 24 business hours. This excludes weekends and holidays.
Each rotary microtome is compatible with low-profile and high-profile blades. With the purchase of a rotary microtome, we will ship a box of 50 low-profile blades free of charge.
Low profile blades are typically used to section thinner slices of paraffin embedded tissue as they are thinner blades. These low profile blades can section as low as 3µm. High profile blades are suitable for thicker slicing of paraffin embedded tissue.
If you are getting thick-and-thin sections while cutting, or you are getting skipped sections, it could be due to several reasons:
When your tissue sections are compressed, wrinkled, or look “scrunched up,” it can be due to:
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