How do you section the brain tissue?

How do you section the brain tissue?

To make uniform, thin sections, the brain must be frozen. A wide variety of sectioning devices are available, each with their own requirements. With a microtome, for example, a very sharp knife is drawn manually across a brain that has been frozen to a chilled platform using dry ice.

How do you embed brain in agarose?

For embedding, we place the rat brain in a 12-well culture dish and fill the well with agarose (once the solution reaches a temperature of around 40 degree C). We then put the dish in a fridge at 4 degree C for about 10 minutes until agarose hardens.

What is a cryostat used for?

Cryostat: A chamber that can maintain very low temperatures. Medical laboratories use a cryostat to preserve frozen tissue samples while a microtome, an extremely sharp cutting instrument mounted inside cryostats, slices the tissue into pieces thin enough to be observed under a microscope.

Which stain is used for brain?

The Golgi staining technique, also called the black reaction after the stain’s color, was developed in the 1870s and 1880s in Italy to make brain cells (neurons) visible under the microscope.

What is brain Lesioning?

Lesioning is when small areas of damage (lesions) are made in your brain. These lesions target cells that control movement and are used to treat movement disorders, such as essential tremor, Parkinson’s disease, and dystonia. In the 1950s and 1960s, lesioning was a common treatment for these disorders.

What is low melting point agarose?

Low Melting (LM) Agarose is a low melting temperature agarose with the highest resolving capacity for large DNA fragments, including PCR products. Low melting temperature allows for the recovery of undamaged nucleic acids below denaturation temperature.

How is tissue sent for frozen section?

During an operation, tissue is transferred to the frozen tissue lab directly from the operating room. There, it is placed on a freezing microtome machine where the bottom of the sample is frozen within seconds.

What is the principle of cryostat?

The cryostat is the instrument to freeze the tissue and also to cut the frozen tissue for microscopic section. The rapid freezing of the tissue sample converts the water into ice. The firm ice within the tissue acts as embedding media to cut the tissue.

What is a cryostat at what temperature range is it operated?

The cryostat can operate between -50°C and + 50°C. The temperature of the thermostated bath (Tc) is measured by a thermocouple with an accuracy of 0.3°C.

Why do neuronal bodies stain blue?

Because ammonium sulfide is a reducing agent, the stain is initially decolorized and the cell bodies appear blue only during fixation and dehydration. This makes differentiation somewhat difficult to control, and after dehydration it may be necessary to return to fixative for further differentiation.