Biomedical research experiments paraffin section technique(5) clinical experimental
The operation of the process of embedding clinical experimental
Embedding, it will have heated the temperature boxes on the stage, remove from the temperature boxes in full bloom of pure paraffin wax cup, poured into the tray with the embedded, remove the storage of materials, wax cup 3, the rapid light Light to clamp the clip to take material flat on the tray at the bottom (note that aspect down place), and then gently warm tweezers toggle materials, to make it neat. Handle gently on both sides of the carton raised, slowly horizon on the basin, and immediately make it sinking into the water, so that box-embedded blocks of the rapid solidification. To be completely solidified paraffin wax (about 30min) can be removed after the backup.
2, slice
will have fixed and repaired wooden platform mounted on paraffin blocks of the clip materials slicer table.
will be fixed on the knife slicing knife clip on the knife-edge upward.
shaking to promote spiral, so that paraffin block with the knife-edge close to, but not more than knife-edge.
paraffin block and knife-edge to adjust the angle and position between the blade and into the paraffin sections of about 15 degrees.
regulator to adjust the thickness of the slice to the desired thickness, generally 4 to 10 microns.
all readjust the emperor can start slicing. At this time shaking his right hand wheel, so that cut into wax with a wax block, left hand holding a brush the wax with a raised, shaking revolution speed is not too fast, usually in the 40 ~ 50r/min.
cut the wax into the 20 ~ 30cm long, with another branch right wax with a brush gently stir to avoid curling and pulling into a belt, flat on the wax with a box on the flank side by a more smooth , down more than wrinkled side facing up.
single-sided blade with a short cut to take wax and put on a glass containing a drop of water placed under the magnifying glass or microscope observation of sections is good.
slice after work should be wiped with chloroform removed a knife slicing knife stained paraffin, wipe clean the slicer for proper preservation.
3, Chip
, take a clean glass slide, drop slide, a drop of adhesive tablets in the central, and then use fingers to smear wash, then into a uniform thin layer.
drop 1 to 2 drops of distilled water has been on a slide coated tablets sticky.
, folder with a small tweezers to take a pre-slit with a blade and wax band on the surface of the water, pay attention to light wax film formed on the side affixed to the slide and make it at the end of Shaopian slide the other end to facilitate pasting labels.
, the film gracefully slide position in the alcohol lamp flame the top of the wax film is heated to a moderate stretch. Or place of exhibition pieces placed in a pre-heating stage (temperature maintained at 40 ~ 45 ). At this point wax heated and stretched because of smoothing. Shaopian make it in one end of the slide, the other end to facilitate pasting labels.
After the slide show on the flat plate on a good mark compiled warm box at 37 drying, a dry day and night, you can remove the boxes deposited in the biopsy to be infected.
10, dewaxing rehydration
Paraffin sections were xylene dewaxing of 5 ~ 10min, and then placed in 100%, 95%, 90%, 80%, 70% alcohol solution at all levels in the 3 ~ 5min, then placed in distilled water in 3min.
Staining the majority of aqueous solution, therefore, must be wax off before dyeing, so that slices of the material from the organic phase into the aqueous phase. Commonly used xylene dewaxing, we should gradually re-wax process of leaching of water and dehydration just the opposite, but because of thin wax film, the time is shorter than the dehydration of multi-dipped wax.
11, dyeing experimental
Slice into hematoxylin staining in about 10 ~ 30min. Staining time should be based on the maturity of stain and the temperature level, appropriate shortened or extended. At room temperature is high for dyeing, dyeing time can be a bit short, otherwise, may be appropriate to extend the time for the winter low temperature can be placed in incubators staining.
12, washed Medical
Rinse with tap water at about 15min. Washing process, to slice the color blue (or into the alkaline water can also be, but the blue-promoting agent in Iraq with the red dye may be refused, if the time in time, should be running water so that the blue chips show is appropriate), but should pay attention to water can not be too large to avoid slicing off, and keep checking with a microscope, see the color blue so far. Tillotts Pharma AG