Impregnation , Embedding , Trimming , Section cutting , Staining & Mounting Flashcards
Give the 3 types of Impregnation
Paraffin wax , Celloidin Impregnation , Gelatin Impregnation
In embedding paraffin embedded tissue are arrange at the bottom of the mold and immerse in melted paraffin on what specific temperature ?
5-10°C above its melting point
a type of impregnation used when dehydration is to be avoided and when tissue are to be subjected to histochemical enzyme study
Gelatin Impregnation
most common simplest and best embedding medium for routine tissue processing
Paraffin wax Impregnation
what are the two methods of Celloiden Impregnation
Wet Celloiden and Dry celloiden method
what are those 3 substitue For Paraffin wax
Paraplast , Ester wax , Water soluble wax
Paraffin oven or incubator temp is
55-60°C
A substitue for paraffin wax whos melting point is 46-48°C
Ester Wax
What are those processes that performs paraffin wax Impregnation and Embedding of Tissue
Manual Processing , Automatic Processing and Vacuum Embedding
what is the common melting point of waxes
45°C to 56°C
a type of impregnation suitable for specimens with large hollow cavities
Celloidin Impregnation
Most commonly used water soluble wax
Carbowax
what is the volume of impregnating medium to the tissue
at least 25 times the volume of tissue
An Elliot Bench Type Processor
Autotechnicon
a process under negative atmospheric pressure use for urgent biopsies for dense and hard fibrous tissue
Vaccum Embedding
Method of celloiden impregnation preferred for processing whole eye sections
Dry Celloiden Method
purified paraffin and synthetic plastic polymers with a melting point of 56-57°C
Paraplast
above melting point of paraffin wax impregnation
2 to 5°C above melting point
Product of paraffin containing rubber
Tissue mat
melting point of paraplast
56-57°C
A Celloidin Impregnation Method for bones , teeth, large brain sections and whole organ
Wet Celloidin Method
An Impregnation type use for large tissue sections of the whole embryo
Celloiden Impregnation
Bioloid is a synthetic wax . true or false . give its use
false it is a semisynthetic wax
for embedding eyes
a synthetic wax
Embeddol
Impregnation is also known as
Infiltration
Celloiden Impregnation is for what type of specimens ?
for large hollow cavities , large tissue section of the whole embryo and for bones and teeth
what are the 6 types of blocking out mold
Leuckhart's Embedding Molds Compound Embedding Molds Plastic Embedding Rings and Base Molds Disposable Embedding Molds Celloidin or Nitrocellulose Method Double Embedding Method
A type of Blocking out mold consist of special stainless steel base mold fitted with plastic embedding ring
Plastic embedding Rings and Base molds
What are the 3 example of Disposable Embedding Molds
Peel Away
Plastic Ice Tray
Paper boats
A type of Blocking out mold that facilitates cutting large blocks of dense firm tissues like the brain
Double Embedding Method
A type of Blocking out mold with series of interlocking plates
Compound Embedding Molds
A type of Blocking out mold where tissues are embedded in shallow tins of enamel pans
Celloidin or Nitrocellulose Method
A type of Blocking out mold consist of two L-shaped strips of metal
Leuckhart’s Embedding Molds
A type of Blocking out mold for embedding hard tissues and whole organs
Celloidin or Nitrocellulose Method
A type of Blocking out mold where tissue are first infiltrated with celloidin and embedded in a paraffin mass
Compound Embedding Method
A type of Microtome Knives use for cutting paraffin embedded on rotary microtome
Biconcave Knife
In microtome knives its an angle formed between the cutting edges
Bevel angle
A type of Microtome Knives use for cutting celloidin embedded tissue block on a sliding microtome
Plane Concave Knife
In Microtome Knives its an angle between the cutting facet and the tissue block
Clearance Angle
Angle of Bevel
27° to 32°
A type of Microtome Knives use for frozen sections or cutting extremely hard and tough specimens embedded in paraffin block using base sledge type or sliding microtome
Plane Wedge Knife
A type of Microtome Knives that has both sides concave
Binconcave knife
A type of Microtome Knives that has both sides straight
Plain wedge knife
What are the two Stages in Sharpening a Knife
- Honing
2. Stropping
what are those three substance use in Honing
Belgium Yellow
Arkansas
Fine Carborundum
A stage in Sharpening Knife that removes burr or irregularities that have been formed during honing
Stropping
What should be the diameter of the wax that surround tissue block
at least 2mm
Other Name for Celloidin Method for Embedding
Nitrocellulose
A stage in Sharpening Knife that removes gross nicks and involve grinding and cutting edge of the knife on a stone
Honing
A substance use for Honing that gives more polishing effect
Arkansas
It is for final polishing of the knife edge
Stropping
Direction use in Stropping
TOE TO HEEL
How many strokes use in Honing
20-30 double strokes
Direction use in Honing
Heel to Toe
How many strokes use in Stropping
40-120 double strokes
What are the 3 types of Microtome Knives
Plane Concave Knife
Biconcave knife
Plane Wedge Knife
Clearance Angle
0-15°
what is the size of the average tissue block
2 to 4 mm thick
Step in Histopath where in tissues are cut into uniformly thin slices with a microtome
Section Cutting
Kind of Microtome that cuts celloidin embedded sections
Sliding Microtome
Kind of Microtome for gelatin and celloiden tissues
Rocking Microtome
Kind of Microtome for general purpose microtomes for cutting semi thin to thin sections for light microscopy
Rotary Microtome
Thickness cut by Rotary Microtome
4-6µ thickness
Kind of Microtome that is dangerous because of the risk of vibration
Sliding Microtome
Thickness cut by Rocking Microtome
10-12µ
Thickness cut by Sliding Microtome
7-9µ
size of sections cut from the block
2-8µ in thickness
Who Invented the Rocking microtome
Padwell Trefall(1789)
Who invented the Sliding Microtome
Adams (1789)
Who invented Rotary Microtome
Minot (1885-1886)
Kind of Microtome that has refrigerated chamber maintained at temperature near -20°C
Freezing Microtome
Paraffin blocks routinely cut at what thickness
5µ
Knive use in Sliding Microtome
Biconcave Knive
Knive use in Rotary Microtome
Plane Concave Knive
Kind of Microtome that cut section for Electrom Microscopy
Ultra Thin Microtome
Who invetned Freezing Microtome
Queckett (1848)
Ultra thin microtome cut tissue at what thickness
0.5µ
In ultra thin microtome tissue is fixed with what fixative agent
Osmium Tetroxide
In freezing microtome temperature are maintain at
-20°C
Enumerate the different kinds of Microtome
Rocking Rotary Sliding Freezing Ultrathin microtome
In Freezing Microtome tissue are cut at what thickness
5-10µ
In Section Cutting It is added to prevent growth of molds
Thymol
What are the 3 types of Tissue Sections
Parrafin , Celloidin , Frozen Sections
Most commonly used Adhesives
Mayer’s Egg Albumin
Type of Tissue Section from tissues that have been fixed and frozen with CO2 or Frozen with Cryostat
Frozen Section
In Celloidin Sections tissue are usually cut at what thickness
10-15µ
In Paraffin Sections tissue are cut at what thickness
4-6µ
Floating-Out Bath Melting Point
6-10°C lower than the wax melting poin
Type of Tissue Section that may be cut by sliding microtome
Celloidin Sections
Type of Tissue Section that may be cut by rocking and rotary microtome
Paraffin Sections
Name the Types of Tissue Section that do not form ribbon
Celloidin Sections , Frozen Sections
Floatin Out Bath is maintained at what temperature
45-50°C
Enumerate 5 Common Adhesives
Mayer's Egg Albumin Dried Albumin Gelatin Starch Paste Plasma
Excess albumin should not be wiped off because it may interfere with staining process hence will take up the stain . True or False
False . It should be wipe off
Steps in Fixing Tissue Sections onto a slide
- Proper use of adhesives
- Proper Orientation
- Deparaffinization
Steps in Fixing Tissue Sections wherein there is a removal of excess wax
Deparaffinization
Steps in Fixing Tissue Sections that promotes adhesions of sections which is spread thinly
Proper use of Adhesives
Steps in Fixing Tissue sections onto a slide wherein tissue is arranged in precise in position on the slide
Proper Orientation
what are the 3 ways in removing excess wax
- Heating of slide over alcohol lamp
- an oven at 60°C
- using xylene
What is the purpose of placing the coverslip over tissue?
to assume permanency of the specimen.
To increase the refraction
What are the Objectives in Mounting
- Protect the specimen from physical injury
- Protect the section from bleaching or deterioration
- Facilitate easy handling storage
- Prevent the damage of sections which may lead to distortion of image during microscopic examination
What are the 2 Techniques in Mounting
A.Slide lowered onto coverslip and quickly inverted
B. Coverslip lowered onto slide
It is a syrupy fluid applied between
the section and coverslip setting section firmly
Mounting Medium
What are the 2 Groups of Mounting Media
Aqueous and Resinous Media
What are the components of Aqueous Media
- Gelatin/Glycerin Jelly/ Gum Arabic
- Glycerol
- Sugar
Group of Mounting Media recommended for majority of staining methods
Resinous Media
What are the 4 characteristics Mounting Medium
The refractive index should be near as possible to that of the glass(1.518) to avoid distortion of image
- Should not Dry Quickly
- Should not dissolve out or fade tissue sections
- Should not cause shrinkage and distortions of tissues
- Should set hard producing
what are the 2 types of Resinous Media
Synthetic and Natural
It is a component of Mounting medium to solidify the media
Gelatin/Glycerin Jelly / Gum Arabic
It is a type of Resinous Media used for embedding undecalcified bones
Synthetic
A component of Mounting Media use to increase the refractive index
Sugar
Groups of Mounting Media use to mount water miscible preparations
Aqeuous
Groups of Mounting Media for prepartion that have been dehydrated and cleared with xylene and toulene
Resinous Media
A component in Mounting Media that prevents cracking and drying
Glycerol
Enumerate the 4 Natural Synthetic Resinous Media
Canada Balsam , DPX , XAM and Clarite
Resinous Media is use on what types of microscope
Electron Microscope and Light Microscope
How many sections in embedding in light microscope
1-2 sections
A type of Natural Synthetic Resinous Media recommended for whole mounts and thick sections
Canada Balsam
A type of Natural Synthetic Resinous Media that is recommended for small tissues drying
DPX
A type of Natural Synthetic Resinous Media that is a mixture in xylene
XAM
Clarite Refractive index is ?
1.544
Dpx refractive index is ?
1.532
XAM refractive index is ?
1.52
Canada Balsam Refractive index is ?
1.524
What are the 3 advantages of Canada Balsam
- Does not shrink much
- Sets Hard without granulation
- Miscible with xylene
What are the 2 advantages of XAM
- Dries Quickly without retraction
* Preserves stain well
What are the advantages of DPX
*Recommended fir small tissues drying
What are the disadvantages of Canada Balsam
Expensive
What are the disadvantages of DPX
- Shrinkage produce
* Not for whole mount
Cover slip size for Large
22/24x56/60 mm
Cover slip size for Medium
22/24x40mm
Cover slip size for Small
22x22mm
What are the 5 types of Aqueous media
Water ,Glycerin , Gum Arabic , Apathy and Bruins
R.I. of Gum Arabic
1.43
R.I. of Apathy medium
1.53
R.I. of Glycerin Jelly
1.44
A type of Aqueous media use for methylene blue nerve preservation
Apathy Medium
A type of Aqueous media use for frozen section
Bruin’s fluid
A type of Aqueous media use as preservative
Glycerin Jelly
A type of Aqueous media use when dehydration and clearing cannot be made
Glycerin Jelly
Gum Arabic is also known as
Farrant’s media