[2S] UNIT 7 Impregnation and Embedding Flashcards
Process of saturating the tissue with a medium, usually liquid paraffin, to permeate or fill up the natural cavities, spaces, and interstices of the tissue.
Impregnation
A suitable embedding mold is filled with the molten wax, the tissue is placed in it and oriented so it is sectioned in the proper plane.
Embedding
T/F: Wax to be used for embedding must contain trace of clearing agent, dust particles, and must be rapidly cooled to reduce the wax crystal size.
F; must not contain trace of clearing agent
A variety of molds can be used depending on the technician’s preference
Embedding
TECHNIQUES FOR EMBEDDING TISSUES
Transfer the tissue with _____ forceps to a small container of freshly melted paraffin
warm
TECHNIQUES FOR EMBEDDING TISSUES
T/F: All parts of the forceps are heated in an alcohol lamp or in a forceps warmer, tips should be hot enough so paraffin does not solidify, but not so hot as to cause paraffin to smoke
F; only the tips are heated
TECHNIQUES FOR EMBEDDING TISSUES
Fill the bottom of the mold with a small amount of paraffin. The depth of the mold should be at least ______ the thickness of the tissue.
twice
TECHNIQUES FOR EMBEDDING TISSUES
T/F: Pick up tissue, and place into the mold. Manipulation of the tissue in the mold must be slow, so paraffin does not begin to harden.
F; must be quick
TECHNIQUES FOR EMBEDDING TISSUES
After tissue is in the mold, fill mold entirely with the paraffin. As the paraffin begins to harden insert a _____ ________ label; the label should not go down to the bottom of the paraffin
code number
TECHNIQUES FOR EMBEDDING TISSUES
After the paraffin block hardens, where do we immerse the mold to hasten solidification of the paraffin?
Into a shallow, cool (10C) water bath, 10-15 mins
TECHNIQUES FOR EMBEDDING TISSUES
T/F: When paraffin is soft, remove it from the mold.
F; When paraffin is completely hardened, remove it from the mold.
T/F: If paraffin is properly cooled, the crystals of paraffin are small and contiguous with each other.
T
Temperature that prevents cracking of the tissue block
10˚C
COOLING TEMP
T/F: The paraffin will appear clear and homogeneous and there is no layering of the paraffin. Paraffin demonstrating these conditions is best for sectioning.
T
Specimen should be embedded and oriented in the center in a manner that it would be surrounded ___ by paraffin on all sides
2 mm
Ideally, the medium should be soluble in processing fluids. It should be easy to section and make ribbons, molten between ______ C
30 to 60 degrees Celsius
Translucent or transparent, stable, homogenous, capable of flattening after ribboning, non-toxic, odorless, easy to handle, and inexpensive
Embedding medium
EMBEDDING MEDIUM
T/F: The properties of the medium should nearly match those of the tissues to be sectioned and should be inert to the embedded material
T
rapidly converted from solid to liquid form on heating
Paraffin
solidifies relatively quickly on cooling
Paraffin
permeates the tissue in a liquid state
Paraffin
becomes fluid on heating to a temperature which will not damage the tissue
Paraffin
when the paraffin solidifies it becomes firm enough to section at _____ _______
room temperature
ADVANTAGES
Time of infiltration and subsequent embedding are relatively short for small pieces of tissue.
Paraffin
ADVANTAGES
Thin sections can be cut with the rotary microtome and sections will adhere to each other to form a ribbon.
Paraffin
ADVANTAGES
Tissue once infiltrated and embedded can be stored in a dry condition indefinitely without damage to the tissue.
Paraffin
- highly purified paraffin waxes with DMSO
(dimethylsulfoxide) - elastic & resilient
Ester Wax
5 Substitute Of Paraffin Waxes
- Paraplast
- Fibrowax
- Bioloid
- Embeddol
- Ester Wax
mixture of highly purified paraffin and synthetic plastic polymers
Paraplast
It is less brittle and less compressible than Paraplast.
Embeddol
is a semisynthetic wax recommended for embedding eyes
Embeddol: Bio/aid
has a lower melting point (46-48°C), but it is harder than paraffin.
Ester Wax
with melting points of 38-42°C or 45-56°C
Water Soluble Waxes
It is not soluble in water, but is soluble in 95% Ethyl Alcohol and other clearing agents
Ester Wax
A polyethylene glycol is suitable for many enzyme histochemical studies. Cytologic details are excellently preserved
Water Soluble Waxes: Carbowax
DISADVANTAGES
Distortion of the histology of the tissue due to shrinkage may occur, especially when sections are being attached to glass slides
Paraffin (paraffin artifact)
Histotechnologist may experience an unpleasant and annoying oyster or garlic taste
Dimethyl sulphoxide (DMSO)
DISADVANTAGES
sectioning of paraffin is difficult at high temperatures.
Paraffin