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What is Disposable Microtome Blades?
Disposable microtome blades are thin, precision-sharpened blades designed for single-use cutting in histology and pathology laboratories. These blades are attached to microtomes, which are instruments used to slice biological tissue into extremely thin sections for examination under a microscope.
There are several options when deciding the best microtome blades for your process. Disposable microtome blades come in different sizes and materials – each designed to slice through various types of material. Your selection relies heavily on the type of microtome equipment you use. To understand more about the specifications of each type of microtome blade, here is a comprehensive guide to the seven most common types of blades used.
1. Rotary microtome blade
Rotary microtomes require a slightly heavier knife, usually measuring between 0.5 to 60 µm. These systems are great for cutting semi-thin to thin sections for light microscopy. They are designed with a hand-wheel operated blade, with the option to modify the angle of the knife to cut larger blocks of tissue as needed.
2. Sledge microtome blade
Sledge microtome blades are significantly larger, measuring around 24cm in length with a wedge shape to minimize vibration. This makes these blades ideal for cutting larger blocks of tissue compared to other machines. A sledge microtome works by setting the tissue block on a steel carriage, which then slides back and forth over a fixed blade. Sledge microtome blades are perfect in applications like segmenting of an entire brain or other large organ tissues.
3. Vibrating microtome blade
A common tool of histochemistry, vibrating microtomes were developed to section fresh plant or animal materials for studying. It operates with a high-speed vibration, aiding in the cutting of soft materials immersed in fluid. This process requires disposable, double-edged razor blades; however, some specialized knives are available.
4. Ultramicrotome blade
Ultramicrotome blades are known for their ability to cut incredibly thin sections. This is perfect for both light and electron microscopy. These blades are made from materials like glass, diamond, or sapphire in order to maintain a very sharp edge for uniform, ultra-thin cuttings.
5. Laser microtome blade
In contrast with the other microtome blades, this process does not utilize a physical blade for cutting sections of tissue. Rather, laser microtomes are built with a bladeless femtosecond laser, to produce samples with great precision. Microtome lasers can produce sections with a thickness of around 5 to 100µm. This method works well on biological materials and a range of other materials.
6. Saw microtome blade
For harder materials, such as bone, ceramic, or resin-embedded samples, saw microtomes are the best option for processing your sample. Materials are carefully cut using a rotating diamond-coated saw. There are some limitations for the sizing of sections, as you will not be able to produce sections smaller than 20µm with this kind of blade.
7. Cryostat blade
Cryostat blades were designed to cut thin sections of frozen tissue. A cryostat is built as a deep-freeze cabinet able to house a rustproof microtome. These systems are compatible with a multitude of blades depending on both the microtome model and the materials that will be cut and sectioned.
Strongly plano-concave: A plano-concave knife has one straight side and a hollow-ground side. This type of profile is extremely sharp and suitable for cutting soft, embedded materials. You'll want to avoid cutting hard materials with a plano-concave knife, which can cause the blade to vibrate.
Biconcave: A biconcave knife has two hollow-ground sides. Like a plano-concave knife, a biconcave blade works best against soft materials and shouldn't be used to cut hard samples.
Plano-concave
A plano-concave knife is similar to the strongly plano-concave profile except it has a thicker back. This blade can be used to cut materials that are too hard for strongly plano-concave blades, and it can also be applied to softer materials, like tissues embedded in paraffin wax. You can find plano-concave knives with various levels of concavity.
Wedge-shaped
Wedge-shaped knives are more rigid than concave knives and can be used to cut harder materials. However, since wedge-shaped knives have thicker tips, they cannot be ground as sharp as concave-profile blades. Nevertheless, wedge-shaped knives can be used to cut paraffin-embedded materials, frozen sections and resin-embedded samples.
Tool-edge shaped
Tool-edge knives are more stable than the other blade profiles but are also the least sharp. Tool-edge knives are commonly used to cut hard materials, synthetic resin blocks and large wax blocks.
Various Disposable Microtome Blades Materials
Steel: Disposable steel microtome blades are made of high-quality stainless steel. When held in an appropriate knife holder, disposable blades produce superior sections. You might use steel blades to cut through plant, and animal and human tissues.
Glass: Glass knives can be used to slice materials for light microscopy and create very thin sections for electron microscopy. Glass knives are hard and extremely sharp, but they're also breakable, so they require careful handling. Since glass knives may deteriorate with storage, they are usually produced immediately before being used in a microtome.
Diamond
Diamond microtome knives are designed to produce very thin slices for light and electron microscopy. Diamond microtome knives are incredibly durable. You can select disposable microtome blades with an amorphous diamond coating to cut through hard tissues and decalcified bone.
Sapphire
Sapphire microtome knives are manufactured from artificially produced sapphire. Sapphire knives have very hard cutting edges and high durability. Sapphire knives can be used with all types of materials, but they require a particular type of knife holder.
Tungsten carbide
Tungsten carbide knives are extremely hard and resistant to wear but are also brittle and must be handled carefully. Tungsten carbide knives may be used to section hard materials, such as resin-embedded tissues.
Non-corrosive
Non-corrosive knives are manufactured from heat-treated stainless steel and contain 12 to 15% chromium. Non-corrosive knives are used for cryostat applications.
Tips For Buying Disposable Microtome Blades
Blade prices vary depending on the material and the distributor. For example, a quality blade will likely cost more than a blade made of laser quality steel. You'll also want to compare prices between distributors to find the right price for your budget.
Make sure to purchase microtome blades from a trustworthy supplier. You want to be able to ask questions if you have any concerns or run into issues, so it's critical to purchase microtome equipment from a company that focuses on customer satisfaction. Ask where its made.
Whenever you buy lab equipment, it's important to investigate the brands and models you're considering. Choose brands with a history of excellence. Ask where the material is sourced from.
A microtome knife is the central component of the machine and not something you want to skimp on when it comes to quality. Make sure to prioritize quality and sharpness when you choose new microtome blades to keep productivity levels high.
You may want to refer to your microtome's manual to ensure you select a knife that will work well with the machine and the knife holder. The manual should tell you the types of knives you can use and the sizes that'll fit your microtome. Or contact your local representative at Mercedes Scientific for top recommendations.
What is Coverglasses?
A cover glass is a thin, flat glass sheet of transparent material, usually square or rectangular, about 20 mm (4/5 inch) wide and a fraction of a millimeter thick, and placed on an object observed with a microscope. The object is usually placed between a cover glass and a slightly thicker microscope slide. The microscope slide is placed on the microscope platform or slide frame and provides physical support for the object and sliding.
What Is The Function Of The Coverglasses?
Protect the specimen
The Coverglasses helps to protect the specimen on the microscope slide from damage, contamination, and evaporation. It creates a sealed environment that helps to preserve the specimen for observation under the microscope.
Prevent distortion
Placing a Coverglasses over the specimen helps to prevent distortion caused by air currents or changes in temperature. It also helps to flatten the specimen, making it easier to focus on and observe under the microscope.
Improve optical clarity
The Coverglasses helps to improve the optical clarity of the specimen by reducing light refraction and reflection at the air-glass interface. This allows for clearer and more detailed observation of the specimen under the microscope.
Thickness Of Coverglasses
Optical Compatibility: Many high-power objective lenses are designed to work optimally with a specific coverglass thickness. For example, standard high-resolution objective lenses such as those used for fluorescence microscopy and other high-magnification applications, are typically corrected for a No. 1.5 coverglass thickness.
Resolution and Clarity: The correct thickness maintains the optical path length that the microscope objective is designed for, ensuring optimal focus and sharpness. Deviations from the intended thickness can degrade image resolution and contrast. This also includes affecting the refractive index and light path, influencing image quality and brightness; providing inaccurate results.
Corrected Aberrations: Using the correct coverglass thickness ensures that spherical aberrations and other optical distortions are minimised.
Sample Protection: Thinner coverglasses are preferred for delicate samples to minimise the weight and pressure on the specimen.
Mounting Media: The thickness of the coverglass should be compatible with the mounting medium and the sample preparation technique used, ensuring that the sample is correctly placed between the slide and the coverglass without air bubbles or gaps.
Live Cell Imaging: For live cell imaging, thinner coverglasses are often used with specialised chambers or dishes to facilitate close proximity of the objective lens to the sample while maintaining the cells in a suitable environment.
These coverslips have a refractive index (1.35 to 1.4) close to water and are made of an acrylate resin (M Brenner et al. 2011). Plastic coverslips are primarily used in education and are not recommended for photomicrography or use with polarized light microscopy because they are birefringent. With fluorescence microscopy some plastic coverslips are auto-fluorescent. Plastic coverslips for general or educational use are lower in cost, unbreakable, and disposable. Coverslips come in different thickness; choosing the correct thickness of coverslip is one of the most important tasks in achieving high resolution and sharp photomicrographs. Using an incorrect thickness coverslip results in spherical aberrations which result in soft hazy images.
Most microscope objectives are designed for use with coverslips that are 0.17 mm thick when the specimen is in direct contact with the overlying coverglass. The 0.17 number is printed on the objective sleeve. Objectives of higher numerical aperture (NA) greater than 0.4 are more sensitive to the thickness of the coverslip than those of lower NA. Objectives starting at 20X NA 0.4 up to NA 0.95 (63X) are most sensitive to spherical aberration caused by incorrect coverslip thickness.
The thickness of 0.17 mm on the objective does not refer to just the coverslip thickness, but the combination of the coverslip thickness and the thickness of medium above the specimen (M. Kozbek 2001). Therefore thick specimens with large amounts of medium above the specimen require thinner coverslips. Experienced microscopists generally choose No. 1.5 coverglasses that are 0.17 mm thick when the specimen is in contact with the coverslip or close to it. Some researchers and technicians grow cells directly on the coverslip and then mount them on a slide. I use No 1.5H coverglasses most of the time but I found that sometimes using thinner No. 1 coverslips are better due to varying amounts of overlying fluid. One way to reduce water or saline above a specimen is to draw the extra water out from below the coverslip by using a small piece of paper towel, filter paper, or allow time for evaporation. This will result in sharper pictures most of the time.
Coverglasses are available in different thickness ranges: Number 0, 1, 1.5, 1.5H, 2, 3, and 4. Higher numbers indicate thicker coverglasses. The proper thickness for most objectives is 0.17mm (some older objectives were made for use with 0.18 mm coverglass). Some companies offer No 1.5H coverglasses with a tolerance of +\- 0.005 mm thick (H stands for high performance). These 1.5H coverslips cost about $100\10,000 but prices from different companies can vary by more than ten fold.
Cover glasses are sold as round, square or rectangular in size depending on their intended use. Rectangular coverglasses are often used for blood smears or on hemocytometer slides, and I found their larger size is useful for scanning plankton. Round coverglasses are easier to “ring” i.e. seal them for long term storage and 22 x 22 mm square coverglasses are commonly used for other specimens.
(1988, 2017) measured the thickness of coverglasses in a package and showed they have a normal distribution around the mean thickness specified. Some researchers measure the thickness of the coverglass with a digital micrometer for critical use. Measuring the coverglass thickness isn’t routinely done as it is time consuming, and the exact amount of fluid above the specimen is difficult to control accurately. However, researchers sometimes measure the coverslips thickness for confocal laser scanning microscopy or when the highest resolution and maximum quality images are required.
Sometimes aquatic subjects I photograph with a microscope are not clear and sharp even when using 1.5H coverglasses and high NA objectives (40X and 63X), but as the water under the coverslip evaporates the coverslip comes into contact with the surface of the aquatic organisms and the image quality improves. I tested No 1 coverslips for live aquatic specimens and found that the images were often clear and crisp.
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