The * step of the slice must be rough cut. The thickness of the rough cut is about 50-100 microns, and the hard tissue or the smaller tissue should be thinner, and the rough cut will be finely cut after the whole tissue is exposed. Finely cut until the surface of the tissue block is uniform. After no white spots, the cut wax pieces are placed in warm water. When slicing, it is required to use uniform force and softness, and the shaking speed is not too fast or too slow. Too fast will result in uneven thickness and machine wear; too slow will thicken the slice. The thickness of the slice is generally 3 to 5 microns. The requirements for slicing are complete, thin and uniform. The size and shape of the cut film should be consistent with the tissue block, and the slice is incomplete, which often misses important lesions, leading to misdiagnosis and missed diagnosis. When slicing a wax block, attention should be paid to the direction of tissue embedding. The direction of the tissue, the direction of the fibers, muscles, etc. should be parallel to the slicing knife. The more difficult part should be placed on top, such as the epidermis of the skin, the envelope of the mass. , the serosa of the gastrointestinal tract, etc., which can reduce the phenomenon of tissue breakage. Apply even force when operating the microtome to avoid excessive force. For decalcified tissue, bone marrow, and known calcified tissue, a fixed knife edge should be used to reduce the possibility of gaps in other areas. When using a brush to stretch the sheet, prevent the filament from entering the knife edge, because each time a pen is cut, a gap is added. The thickness of the slice is generally 4 to 6 micrometers. Some people think that as long as the microtome is marked with a few micrometers, the sliced ​​slice is a few micrometers. In fact, when the slicing knife is not sharp, or the speed is not uniform when slicing, or when the cutting is slow, the cut film will become thicker. Only when the microtome is very sharp and sliced ​​at a constant speed can the thickness be truly guaranteed. The following are the causes of the problems encountered during slicing and the possible treatment methods: (1) the tissue is brittle: generally dehydrated, transparent, waxing time is too long, the temperature is too high, and is also related to the texture of the tissue itself, when slicing It may be better to blow the wax sheet with your mouth while cutting. (2) The slice is rolled up, it may be that the knife is not sharp, or the blade is on the other side, or the knife angle is too large, the slice is too thick, and so on. (3) Wafer bending: It may be that the blade is uneven, the slicing knife is not straight, and the slicing knife is not parallel to the wax block. (4)
Transparent, waxing time is too long, temperature is too high, and it is also related to the texture of the tissue itself. (5) Uneven thickness: It may be that the knife, the knife seat and the wax block are not clamped, the tissue is too hard, or the slicer spindle is too forward. , or the slicer has worn out. (6) Cracks in the slice: there may be a gap in the knife, impurities in the paraffin, calcification, bone fragments or wire knots in the tissue.
There may also be cotton paper fibers and the like. (7) The slice is not contiguous, the slice is not cut, the slice is very thick, or the wax block is white after being sliced, and the invagination: the tissue is poorly dehydrated. Remedy: The wax block can be dissolved first, the tissue is taken out, placed in acetone in a heated water bath (80 ° C), 30 to 60 minutes, and then embedded in a transparent dip wax, perhaps better.
Dehydration and leaf slice dehydration should be from low concentration of alcohol to high concentration of alcohol, 80% alcohol, 1 channel, 95% alcohol 2, pure alcohol 2, (n-butanol 1), xylene 2 channels. Alcohol with a lower concentration is more likely to dehydrate than a higher concentration of alcohol, but it is also easy to discolor the red, so the dehydration time can be shorter, 3-5 minutes per lane, and pure alcohol for 10 minutes per lane. In order to increase the compatibility of alcohol and xylene, a n-butanol can be added in front of xylene; stone carbonate-xylene solution also has this effect, but if it is not washed with stone carbonic acid, it can cause discoloration of the slice, which is not conducive to slicing. Long time, so do not recommend. After the sections were transparent to xylene, they were sealed with a neutral gum. To increase the transparency of the slices, prevent the cells from shrinking, cracking or slicing black crystals. Therefore, we stipulate that the slices must be wet-sealed, and should not be dried in a thermostat or blown dry with a hair dryer to dry the seal. In the winter, spring and mildew seasons in the south, the gas exhaled by the nose and mouth should be prevented from coming into contact with the slice during the filming; in the hot weather, it is not advisable to take out multiple slices at a time to be sealed, so as to avoid the “sweet†and transparent formation. Poor, there are cloud-like drops of water. The replacement of the dehydrating agent should also be stipulated for a certain period of time. Generally, it is suitable to replace 500 pieces of liquid per 500ml of liquid. The sealant gum should not be too thin. When the film is sealed, the gum should be evenly filled with the cover slip and the gum should not be spilled. To cover the entire organization, there must be no air bubbles. After zui, paste the label, the label must be attached to the left side of the slide, the number is clearly written, and the Zui can print.
Transparent, waxing time is too long, temperature is too high, and it is also related to the texture of the tissue itself. (5) Uneven thickness: It may be that the knife, the knife seat and the wax block are not clamped, the tissue is too hard, or the slicer spindle is too forward. , or the slicer has worn out. (6) Cracks in the slice: there may be a gap in the knife, impurities in the paraffin, calcification, bone fragments or wire knots in the tissue.
There may also be cotton paper fibers and the like. (7) The slice is not contiguous, the slice is not cut, the slice is very thick, or the wax block is white after being sliced, and the invagination: the tissue is poorly dehydrated. Remedy: The wax block can be dissolved first, the tissue is taken out, placed in acetone in a heated water bath (80 ° C), 30 to 60 minutes, and then embedded in a transparent dip wax, perhaps better.
Dehydration and leaf slice dehydration should be from low concentration of alcohol to high concentration of alcohol, 80% alcohol, 1 channel, 95% alcohol 2, pure alcohol 2, (n-butanol 1), xylene 2 channels. Alcohol with a lower concentration is more likely to dehydrate than a higher concentration of alcohol, but it is also easy to discolor the red, so the dehydration time can be shorter, 3-5 minutes per lane, and pure alcohol for 10 minutes per lane. In order to increase the compatibility of alcohol and xylene, a n-butanol can be added in front of xylene; stone carbonate-xylene solution also has this effect, but if it is not washed with stone carbonic acid, it can cause discoloration of the slice, which is not conducive to slicing. Long time, so do not recommend. After the sections were transparent to xylene, they were sealed with a neutral gum. To increase the transparency of the slices, prevent the cells from shrinking, cracking or slicing black crystals. Therefore, we stipulate that the slices must be wet-sealed, and should not be dried in a thermostat or blown dry with a hair dryer to dry the seal. In the winter, spring and mildew seasons in the south, the gas exhaled by the nose and mouth should be prevented from coming into contact with the slice during the filming; in the hot weather, it is not advisable to take out multiple slices at a time to be sealed, so as to avoid the “sweet†and transparent formation. Poor, there are cloud-like drops of water. The replacement of the dehydrating agent should also be stipulated for a certain period of time. Generally, it is suitable to replace 500 pieces of liquid per 500ml of liquid. The sealant gum should not be too thin. When the film is sealed, the gum should be evenly filled with the cover slip and the gum should not be spilled. To cover the entire organization, there must be no air bubbles. After zui, paste the label, the label must be attached to the left side of the slide, the number is clearly written, and the Zui can print.
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