Wax Infiltration Protocol

• After the tissue is completely transparent with the transparent agent, it is placed in the paraffin wax which is melted at about 65°C. The process of impregnation in the electric thermostat at 65°C is called wax immersion.
• When the tissue is immersed in wax, the paraffin wax enters all corners of the tissue and is preserved in all corners due to the action of the inducer (xylene). When the paraffin is embedded and cooled, the paraffin immersed in it can play a supporting role, and prevent the tissue from deformation, collapse and other phenomena, so that the section can be completely cut out for easy observation under the mirror.

Improvement of Wax Properties

• Wax heating and cooling method. The wax is placed in a container and heated on an electric furnace. With the extension of time and the increase of temperature, the sound will gradually decrease until it disappears. At this time, the container of wax can be immersed in cold water, wax condensation, and then continue to heat, so repeated several times that melting-cooling -melting-cooling. This treatment method can increase the density and plasticity of paraffin and remove impurities.
• Increasing and improving the toughness and viscosity of wax. This method requires the addition of some things to obtain, such as beeswax and hard fat, the highest melting point of beeswax can reach more than 70°C, increasing it can increase the hardness of paraffin wax.

Traditional Wax Dip Method

• This is a manual wax dip method without vacuum negative pressure.
• According to the melting point of paraffin, determine the temperature of the wax dipping box. If the melting point of hard wax is 62°C, the temperature of the wax dipping box should be 2-3°C higher than the melting point of paraffin, that is, the wax dipping box is adjusted at 65°C.
• The tissue into the soft wax, and then into the hard wax, according to the different tissues, the size of the tissue to determine the wax immersion time, small tissue 20-30 minutes, large tissue several hours.

Vacuum Negative Pressure Wax Immersion Method

Vacuum negative pressure wax immersion method uses a vacuum pump to pull out the air in the box, to form vacuum negative pressure. Under this condition, due to the weightlessness, various molecular activities are accelerated, which promotes the rapid completion of the wax immersion process.

• Increasing the hardness of paraffin wax can also be added fat wax acid, tar, bayberry wax, etc.
• The temperature of the vacuum constant temperature drying oven should be slightly higher than that of the traditional wax immersion method, because when the air is pumped to form a vacuum, the hot air in the box will be extracted, resulting in the temperature in the box falling, so that the melted paraffin condensing into blocks.

Whole Slide Imaging

Transmission Electron Microscopy (TEM) Service