What is the importance of the technique of microtomy?

Important in science, microtomes are used in microscopy, allowing for the preparation of samples for observation under transmitted light or electron radiation. Microtomes use steel, glass or diamond blades depending upon the specimen being sliced and the desired thickness of the sections being cut.

What is the most important step in tissue embedding?

Correct orientation of tissue in a mould is the most important step in embedding. Incorrect placement of tissues may result in diagnostically important tissue elements being missed or damaged during microtomy.

What is the importance of water bath in microtomy?

A flotation bath or water bath is the intermediate step between cutting paraffin sections and placing them on slides. Simply sticking paraffin ribbons on slides will not work! A warm water bath allows tissue to relax and smooth out prior to being mounted on a glass slide.

What are common problems during microtomy?

Microtomy Troubleshooting Tips

• The Ribbon is Not Forming. The most common reasons for the ribbon not forming are either the wax is too hard, or the knife is at the wrong clearance angle.
• Compression.
• Ribbon Comes Off in a C.
• Sections Roll Up on Knife Edge.
• Thick/Thin Sections.
• Knife Line.
• Tears.

What is the other equipment used during microtomy?

Most microtomy is performed on paraffin wax embedded tissue blocks. The basic instrument used in microtomy is the microtome; an advancing mechanism moves the object (paraffin block) for a predetermined distance until it is in contact with the cutting tool (knife or blade).

Who performs microtomy?

As one of the most essential skills a histotechnologist can acquire, microtomy bridges the process between specimen collection and microscopic analysis. The ultimate goal of microtomy in a histology laboratory is to produce slides that are free of artifacts and errors.

What are the factors that influence staining process?

Factors that affect staining include: Concentration of the Dye – The greater the concentration of the dye, the more the dye is bound to tissue components. Temperature – An increase in temperature increases the rate at which the dye diffuses throughout the tissue sample.

What are the factors affecting fixation?

Factors affecting fixation

• Temperature: In general, an increase in temperature increased the rate of fixation but also increased the rate of autolysis and diffusion of cellular elements.
• Size: 1–4 mm thickness.
• Volume ratio: At least 15-20 times greater than tissue volume.
• Time: 24 – 48 hours.

What is the purpose of keeping the block cool during Microtomy?

To promote good ribbon generation, the paraffin block needs to be sufficiently hard to enable the sections to be cut on the microtome. This is routinely achieved by placing the paraffin blocks on ice, cooling both the tissue and the wax to a similar consistency and swelling the tissue to make it easier to section.

What is the other equipment used during Microtomy?

What are the different faults that can be observe during sectioning?

Common Problems with Sectioning

• Cut on an angle. It is important to make sectioning cuts that are perpendicular to the longitudinal axis of the root.
• Cut too thin.
• Cut too thick.
• Dull Razorblade.
• Sections cut in half.
• Exposure to Excessive Fixative.
• Lack of Exposure to Fixative.
• Tissue Too Young.

What are the steps involved in tissue processing?

Overview of the steps in tissue processing for paraffin sections

• Obtaining a fresh specimen. Fresh tissue specimens will come from various sources.
• Fixation. The specimen is placed in a liquid fixing agent (fixative) such as formaldehyde solution (formalin).
• Dehydration.
• Clearing.
• Wax infiltration.
• Embedding or blocking out.

How to prepare the microtome for section cutting?

Accordingly, the microtome was prepared for the process of section cutting: The indicator for the thickness of the section should be adjusted and kept between 4-6μ. If there is enough paraffin in front of the tissue untrimmed, then the block must be cut until the tissue reaches the knife.

What are the limitations of microtomy?

Careless embedding can make microtomy much more difficult. » Avoid under‑filling the cassette as this can allow unstable clamping in the microtome and lead to cutting “thick then thin” sections and other problems. » Avoid over‑filling cassettes as this can interfere with the correct alignment of the block face for sectioning.

What is the thickness of tissue during microtomy?

INTRODUCTION TO MICROTOMES Microtome is an instrument with the help of which sections of tissues are cut and the process of cutting thin sections is known as Microtomy. The thickness of sections produced during microtomy may be between fractions of 50-100 nm, in ultramicrotomy, to several 100 microns.

What is the most commonly used microtome?

Nowadays, the most commonly used type of microtome is a Rotary microtome which is easy to operate and ideal for routine use for diagnosis and research purposes. ⇒ It is used for slicing paraffin tissue sections of uniform thickness.