The use of the cryostat microtome, a cutting-edge technique, has completely changed how scientists and researchers examine biological tissues. The fabrication of incredibly thin chunks of frozen tissue samples for microscopic examination is possible thanks to this sophisticated technique for tissue sectioning. They have evolved into essential tools in a variety of disciplines, including pathology, pharmacology, and neurology because they can accurately and precisely slice tissues. This article will examine the science of this tool and how they are used in scientific investigations. We will also go through the benefits of employing this technology and how it can help us understand biological processes and diseases better. Take a look at the electric handpiece.
What is a Cryostat Microtome?
A cryostat microtome is a unique laboratory tool used to slice frozen tissues into incredibly thin pieces for examination under a microscope. It makes use of a microtome knife, which slides across the material to cut tissue into precise slices with thicknesses typically between 1 and 20 microns. To avoid stretching or harming the tissue, the samples are kept frozen while being cut. To investigate the cellular and molecular makeup of tissues, cryomicrotomes are frequently employed in the domains of neurology, pathology, and pharmacology. They have transformed how scientists examine biological samples and made substantial strides in scientific inquiry possible. Another microtome you should read about is the rotary microtome.
Parts of a Cryostat Microtome
- Sample holder- It holds the frozen tissue sample in a fixed position for sectioning.
- Microtome knife- It is a sharp cutting edge that slices frozen tissue samples into your desired measurement.
- Microtome chuck- It controls the movement of the knife for precise sectioning
- Cryostat chamber- It maintains the frozen temperature of the tissue for sectioning
- Blade holder- It holds the knife in position and allows it to be tilted to the right angle for slicing tissue samples.
- Section transfer system- This system is used to gather the cryostat microtome-generated sections and convey them to slides or other collection media for subsequent processing.
- Specimen cooling system- This device keeps the sample at a constant low temperature to prevent melting by circulating a refrigerant.
- Waste container- This retains the water produced by the ongoing processes of defrosting and freezing inside the cryostat.
- Anti-roll plate- inhibits the tissue portions from curling after being sectioned by the blade.
- U V source– The cryostat chamber is sterilized using UV light because it effectively cleans surfaces. Generally, disinfection can be accomplished with a 35-minute U.V. exposure.
- Switch- It is used for lighting: used to turn on/off the cryostat.
- UV switch– To turn on the UV when sterilization is required, use the V On/Off switch.
- Defrosting button- It is used to regulate manual defrosting. Once the experiment is finished, the cryostat defrosts.
- Control panel lock- It guards against unintentional changes to the device’s protocol set.
- Timer- The cryostat can be set to operate for a predetermined period of time. It also controls the timing of the defrost.
- Temperature regulation- It regulates the temperature Controller: It moves the tissue sample in the direction of the knife.
What is the principle of the Cryostat Microtome?
The cryostat microtome works on the basis of the tissue sample being frozen to an extremely low temperature and then being cut into thin slices with a microtome.
Using a cryostat, the tissue sample is frozen to a temperature below its freezing point to begin the cryomicrotome’s operation. The tissue sample is then fixed on a sample holder and set on a microtome stage. As the blade advances through the frozen tissue sample, it cuts thin slices. It is possible to precisely manage the section thickness by adjusting the motion of the blade.
Applications for the slices created by the microtome include microscopy and research on tissue structure, biochemical composition, and molecular interactions. Since the freezing procedure lessens tissue damage and deformation, using this tool is especially helpful for maintaining the tissue’s original state.
A step-by-step guide to using the Cryostat Microtome
- Freeze the tissue sample- The tissue sample is subsequently placed in a cryostat and frozen to a temperature below its freezing point. This procedure involves putting the sample in a chamber that is rapidly and uniformly frozen using a coolant.
- Fix the specimen- The specimen is fixed into the sample holder to hold it firmly for sectioning.
- Adjust the blade- Press the buttons or rotate the knobs to alter the position of the blade.
- Section the tissue- Control the blade to section the tissue to your desired thickness. The thickness can be adjusted by the movement of the blade.
- Gather the tissues- The sections are gathered onto a glass slide or another suitable surface, and they are typically kept there until they are needed again.
Benefits
- Preservation of tissue content and structure- The use of freezing preserves the specimen’s original state, causing the least amount of tissue damage and deformation. This makes it simpler to perform an accurate and trustworthy analysis of the tissue sample.
- Accurate section thickness- The ability to precisely control section thickness enables scientists to produce uniform sections with high levels of accuracy.
- Versatility– sections can be utilized for a variety of tasks, such as microscopy, biochemical tests, and molecular analysis due to its adaptability.
- Speed– The use of microtome avoids stages like fixing, dehydration, clearing and embedding.
- Easy to use– It has a user-friendly procedure that is simple enough for researchers to pick up fast with little training. Do not forget about the sliding microtome.
How Should A cryostat Microtome Be Cleaned and Maintained?
For a cryostat microtome to operate at its best, it must be regularly cleaned and maintained. The chamber, blade holder, and other components must typically be cleaned frequently, and the moving parts must be lubricated. In order to properly clean, maintain, and troubleshoot your cryostat microtome model, it is important to follow the manufacturer’s protocol and recommendations.
Conclusion
Researchers can prepare thin sections of frozen tissue samples for observation using this tool, a potent tool for research. The basic idea behind a cryostat microtome is to freeze the tissue sample to a low temperature before using a microtome to slice it into thin layers. The preservation of tissue content, accurate section thickness, adaptability, speed, and simplicity of use are just a few advantages of this method. Researchers have access to trustworthy and precise data with the cryostat microtome that can be used to improve our understanding of biological systems and diseases. Do not forget about the tissue processor.