MOUNTING/LABELING Flashcards
last step on the tissue processing
Mounting
syrupy fluid applied between the section and the coverslip IMMEDIATELY after staining
Mounting medium
Mounting a tissue specimen is essential for preserving the specimen during ______ (for long periods of time) as well as for enhancing _____ quality during microscopy
storage
image
Histopath slides → can last up to _____ years retention period
10
Mounting medium:
[Action] tissue section. Not ____ or blurred
flatten
wavy
blurred
If an unmounted stained section is seen in the
microscope, differences in ______ between the glass slides and the tissue components, may lead to difference in _________; hence, very little microscopic detail can usually be appreciated.
◆ Some parts of tissue sections are [2]
light refraction
length dispersion
blurry/wavy
We need to use _______ to secure properly the glass slide and to prevent movement and washing out or scratches
mounting medium
Mounting medium flatten out sections for what purpose?
fine details of cells
Mounting medium purposes [4]
promote spx adhesion
promote glass slide adhesion
prevent spx movement
spx preserve
Characteristics of a Good Mounting medium
- freely miscible w/ xylene/toluene
- should not dry quickly
- should not dissolve/fade out tissue sections
- should not crack or produce artefactual granularity on the slide upon drying
Dryness will cause _____ on glass slides, resulting to the tendency of tissue section to be destroyed.
cracks
artefactual granularity causes ?
bubbles
No bubbles or cracks must be produced
because you will not appreciate the cells’ [2]
nucleus
cytoplasm
Ratio of speed of light in air and in a specific medium
Refractive Index
Value to which light can pass through a
medium
RI
Refractive index should be as near as possible to
that of the glass which is [#]
1.518
As much as possible the RI must be higher
than the RI of ____ to consider that the mounting medium is of good quality
glass
________
They are designed to mount water miscible preparations directly from water where the stained is removed or decolorized with ____ or _____.
Because there are components of tissue slides that are dissolved when using alcohol or xylene: ______
Aqueous Media
alcohol
xylene
lipids
Aqueous Media is made up of? [4] [+function]
Gelatin, Glycerin Jelly or Gum Arabic- solidifigcation
Glycerol- prevent cracking/drying
Sugar- increase RI
Merthiolate- preservative
Aqueous Media examples [6]
Water
Mineral Oil
Glycerin
Levulose
Brun’s Fluid
Farrant’s medium
Von Apathy’s Gum Syrup
Karo Corn Syrup
Has low RI
➔ Moderately transparent
➔ Evaporates easily
Water
➔ Good only for temporary mounting
Water
May be used also as preservative for Aqueous Media other than merthiolate
Glycerin
➔ Has high refractive index, RI 1.46
➔ Last for a few minutes
Glycerin
Glycerin Jelly RI:
1.47
Farrant’s medium RI:
1.43
Useful temporary mounting media for Romanowsky stained smear
Mineral Oil
Von Apathy’s Gum Syrup RI:
1.52
Von Apathy’s Gum Syrup is made up of? [5]
Distilled water
Sucrose
Pure gum arabic
Pure cane sugar
Thymol
Levulose/Fructose RI
1.5
Brun’s Fluid is made up of? [4]
Distilled water
Glucose
Glyrcerin
Spirit of Camphor
For preparations that have been dehydrated and cleared with xylene or toluene
Resinous media
Recommended for majority of staining methods
Resinous media
Resinoud media is divided by [2]
Natural type of resins
Synthetic type of resins
Canada Balsam
Natural resin extracted from the ______ Tree → ________.
Canadian
Abus Balmasea
type of resin that is transparent/colorless
Canada Balsam
type of resin that sets to a hard consistency when dry
Canada Balsam
There are two categories of mounting media
Organic/Aqueous
Hydrophobic/Hydrophilic
________can only be used for enzymatic labels where the precipitate formed between the enzyme and the chromogen is not soluble in the organic solvents used during mounting of the tissue
Examples:
Organic mounting media
Diaminobenzidine
DAB
Aqueous mounting media is generally suitable for all _______ label/______ combinations and _____ labels.
enzymatic labels
chromogen combinations
fluorescent labels
Synthetic Brands Superior than Canada Balsam [5]
- DPX of Kirkpatrick & Lendrum
- Clarite X
- Permount, Clearmount, Histosec, Harleco synthetic resin, Entallan
- XAM
- Eukitt
meaning of DPX
Dibutyl Phthalate & Xyelene
DPX of Kirkpatrick & Lendrum is also called as
Resin “distrene 80”
Tricresyl phosphate/ Dibutyl phthalate
a good mounting medium but shrink somewhat on drying
DPX of Kirkpatrick & Lendrum
DPX of Kirkpatrick & Lendrum RI:
1.52
When mounting medium has ____, you can see
in the ____ of glass slides, the mounting medium might shrink out
dried
edges
One of the widely used mounting media
Clarite X
Clarite X RI:
1.544
Generally preferred as compared to DPX
Clarite X
XAM RI:
1.52
Synthetic resin mix in xylene, available in [color] (disadvantage) or [color] solution
XAM
pale yellow
colorless
Dries quickly without retraction, and preserves the. stains well
XAM
It is an adhesive and specimen preservative
Eukitt
Eukitt contains [2]
5% Acrylic resin
55% Xylene
In lab we used _____ to mount the slides
Eukitt
Eukitt Properties:
◆ ______ drying with minimal ______.
◆ RI close to the glass (______)
◆ ______ neutral, little sensitivity to _____
◆ No reaction to _____. (Does not change
_____/_____)
◆ Non-______, unaffected by [hot/cold] and ____.
◆ Stays optically clear for over _____ years
◆ Does not _____.
Fast
air bubbles
1.510
chemically
water
staining
color/pH
yellowing
cold
UV
10
crystallize
Finished slides easily reworked by immersing previously mounted slides in ______ to remove coverslip.
xylene
In histopath if you have to remove coverslip you have to dip in _____ or _______ that is miscible to mounting
saline
clearing agent
The slides in mounting media may then be incubated at ___*C for _____ hrs. After mounting, to harden the medium.
37C
12-24 hrs [half-whole day]
Setting may be hastened in a ______ at 50*C for ___ hrs.
hot oven
2 hrs
After _______ staining, mounting media are used to adhere a coverslip to a tissue section or cell smear.
immunochemical staining
Mounting media also play an important role in
improving the _____ and ____ of an image during microscopy.
clarity
contrast
Sealing the margin of the coverslip
Ringing
Aside from putting mounting medium you can _____
media to seal the glass slide and cover slip
pit
a ringing agent and is made of [2 parts]
Krognig cements
paraffin wax
4-9 parts of powdered colophonium resin
The stained section on the slide is covered with a thin piece of glass to protect the tissue from scratches/ damages, to provide better optical quality for viewing under the microscope, and to
preserve the tissue section for years to come.
Coverslipping
With used of coverslip it can preserve the _____
ot tissue it can protect from scratches and to preserve long period of time for years cover slip is used to preserve the section
integrity
Cover slip are identified by ______ which pertain to the thickness
number
Thickness and transparency of the cover slip is [directly/inversely proportional]
As thickness increases, transparency is [increased/reduced].
inversely proportional
reduced
Cover glasses come in different sizes, namely:
[4]
18x18mm
20x20mm
22x22mm
25x25mm
The choice of cover glass depends on the ____ of the tissue to be mounted.
size
Slide with frosted edge, use ________to label the specimen
Slide with non-frosted edge, use ________to label the specimen
LED pencil
Diamond pencil
The labels may be [3]
on each
written
etched
embossed
