HISTOPATH GREGORIOS 40-61 Flashcards
(211 cards)
1
Q
Immediate diagnosis is accomplished through the use of a
A
frozen section
2
Q
It is especially recommended when lipids and nervous tissue elements are to be demonstrated
A
Frozen section
3
Q
Frozen section is especially recommended when __________ tissue elements are to be demonstrated
A
lipids and nervous
4
Q
Frozen sections are usually done on __________ as well as on __________
A
muscle and nerve biopsies
surgically removed tumors
5
Q
A fresh tissue is frozen on a:
A
microtome with co2 / cryostat (-10 to -20 deg)
6
Q
What fixative is used sa frozen section
A
liquid fixative
7
Q
The morphological detail and resolution of frozen sections are [mas maganda/mas panget] compared to that embedded in paraffin
A
MAS PANGET
8
Q
The advantage of the frozen section method
A
Rapid processing time
Less equipment
Less need for ventilation
9
Q
Applications of frozen sections:
A
Rapid diagnosis during sx
Enzyme histochemistry
Demonstration of soluble substances such as lipids and carbohydrates
Immunofluorescent and immunohistochemical staining
Silver stains, particularly in neuropathology
[REDIS]
10
Q
Slow freezing can cause distortion of tissue due to
A
ice crystal artifacts / freeze artifacts
11
Q
The more commonly used methods of freezing include:
A
Liquid nitrogen
Isopentane cooled by liquid nitrogen
Carbon dioxide gas
Aerosol sprays
[LICA]
12
Q
The most rapid of the commonly available freezing agents
A
Liquid nitrogen
13
Q
Liquid nitrogen is generally used in ______ and during ______ procedures
A
histochemistry
intraoperative
14
Q
Main disadvantage of liquid nitrogen
A
liable to crack due to the rapid expansion of the ice within the tissue
overcools urgent biopsy blocks causing damage to both block and blade if sectioning is done at -70°C or below
15
Q
The tissue snap-frozen in
liquid nitrogen must therefore be _____ before sectioning is attempted
A
allowed to equilibrate to cryostat chamber temperature
16
Q
The majority of non-fatty unfixed
tissues are sectioned well at temperatures between:
A
-10°C and -25°C
17
Q
One problem with the use of liquid nitrogen is that it causes a _____
A
vapor phase to form around the tissue
18
Q
Effect of vapor phase
formation around the tissue [liquid nitrogen]
A
acting as an insulator that causes uneven cooling of tissue [muscle biopsy]
19
Q
Ano gagawin pag may vapor phase
formation around the tissue due to liquid nitrogen?
A
freeze the tissue in:
Isopentane,
OCT, or
Freon 2.2
[bec of their high thermal conductivity]
20
Q
Isopentane is [solid/liquid] at room temperature
A
LIQUID
21
Q
A Pyrex glass beaker containing
isopentane is usually suspended in a flask of liquid nitrogen until _____
A
half-liquid and half-solid stage is reached
22
Q
_____ is usually suspended in a flask of liquid nitrogen until half-liquid
and half-solid stage is reached
A
A Pyrex glass beaker containing
isopentane
23
Q
The Pyrex beaker is removed from the liquid nitrogen when _____
A
small crystals start forming on the side of the beaker (approximately -170°C)
24
Q
The beaker is removed from the liquid nitrogen when small crystals start forming on the side of the beaker (approximately -170°C) and the tissue to be frozen is _____
A
dropped into the cooled liquid isopentane
25
Tissue to be frozen in ispentane is affixed on a:
cork disc,
aluminum foil, or
cryostat chuck
26
is adequate for freezing small pieces of tissue except muscle
aerosol sprays
27
_____ have a distinct advantage of rapidly freezing blocks of any type of tissue
Quickfreezing spray cans of fluorinated hydrocarbons (e.g., Cryokwik)
28
Fresh, completely unfixed tissues, or tissues that have been briefly treated
with formalin may not require embedding anymore; instead they may be _____
frozen and cut in a freezing microtome or cryostat.
29
Two methods of preparing frozen
sections may be resorted to:
1. _____
2. _____
1. Cold Knife procedure
2. Cryostat procedure (Cold Microtome)
30
[COLD KNIFE PROCEDURE]
A piece of filter paper soaked in __[1]__ is placed on the microtome
stage, and short bursts of __[2]__ are applied, freezing the filter paper to the stage
1. gum syrup
2. co2
31
Thickness of tissue section sa COLD KNIFE procedure
3-5mm
32
Gano katagal yung short bursts ng CO2 sa cold knife procedure
1-2 seconds duration
4 second interval
33
the point at which sections may then be cut at 10 µm thickness
DewLine
34
[COLD KNIFE PROCEDURE]
The tissue is lifted up to the knife manually and trimmed until _____
the surface is flat
35
[COLD KNIFE PROCEDURE]
After frozen tissue surface becomes flat, it is warmed with the finger until ______
the hard frozen tissue starts to thaw and becomes visible to the naked eye
36
Cold knife microtomy sections do not form ribbons but rather ______
stick to the knife blade
37
Cold knife sections that are stuck to the knife blade must be removed with _____
a camel hair brush or finger moistened with water
38
Cold knife sections are then transferred to a dish of ______
distilled water to separate
39
[COLD KNIFE MICROTOME]
The water dish is usually placed on a
______ background
dark or black
40
[COLD KNIFE]
The surface of the block may then be softened by warming slightly
with the ______
ball of the finger or thumb
41
[COLD KNIFE]
Tissues that have not been sufficiently
frozen will cut __[1]__, and the block may __[2]__
1. thick and crumble
2. come away from the stage
42
Optimum temperature used for sectioning in COLD KNIFE microtomy
Knife = -40° to - 60°C
Tissue = -5° to - 10°C
Environment = 0° to - 10°C
43
[COLD KNIFE]
Sections thinner than _____ generally cannot be obtained even from tissues that section well, and with ideal conditions for sectioning
6 µ
44
The success of this procedure [COLD KNIFE MICROTOMY] depends upon _____
ambient temperature and
humidity
45
Microtome, knife, specimen and atmosphere are kept at the same
temperature
Cryostat
46
Microtome, knife, specimen and atmosphere are not kept at the same
temperature
Cold knife / Cold microtome
47
The cryostat consists of an insulated microtome housed in an ___[1]___ and maintained at temperatures near ___[2]___
1. electrically driven refrigerated chamber
2. -20°C
48
The optimum working temperature of cryostat
-18 to -20°C
49
Majority of the sections can be cut in ___[1]___ conditions, where the
temperature for sectioning can be ___[2]___
1. isothermic
2. accurately established and controlled
50
Fresh frozen tissue requires that the tissue be maintained in the _____ during cutting of section
frozen solid state
51
[CRYOSTAT]
The tissue must be sufficiently cold to prevent ______of cell and tissue structures as the knife passes thru it.
compression and displacement
52
[cryostat]
The microtome knife needs to be chilled and maintained at low temperature to prevent ______
complete melting of the tissue
53
[cryostat]
When the tissue is too cold, on the other
hand, ______ is increased
resistance to cutting
54
The cryostat should be left on at all
times even when not in use, since it will ______
require several hours to reach
operating temperature from a room temperature start
55
It takes at least ______
for a knife to come down to operating temperature, so that a spare knife should always be kept inside the cryostat cabinet
one hour
56
What must be kept scrupulously clean and dry to ensure that the sections will cut smoothly and freely onto the knife surface? [cryostat]
Knife
Undersurface and edge of the anti-roll
57
May be used to clean the knife and anti-roll plate [cryostat]
Soft tissue paper (either dry or moistened with absolute alcohol)
58
The cryostat should be defrosted during the ___[1]___, including cleaning and oiling of microtome with ___[2]___
1. weekend
2. special low temp oil
59
[cryostat]
Require much lower temperatures to impart a suitable consistency for cutting
Fat or mucin
Hard or dense structures in a soft matrix
60
How to section:
Fat or mucin
Hard or dense structures in a soft matrix
Lowering the tissue or knife temperature or both
61
How to lower temperature of tissue/knife in cryostat for sectioning of:
Fat or mucin
Hard or dense structures in a soft matrix
Placing the block holder in a bath of alcohol or acetone containing dry ice,
Exposing the tissue to carbon dioxide
62
______ are generally used as mounting
media for tissue blocks that need to be sectioned on a cryostat
Synthetic water-soluble glycols and resins
63
Recommended mounting media for cryostat sectioned tissues
O.C.T. (Optimal Cutting Temperature) compound, Lab-Tek Products, Division of Miles Laboratories
64
Temperature of mounting media for cryostat
-5 to -15°C
-15 to -25°C
-35°C
65
Cryostat mounting media under -5 to -15°C
[BULL SSCT]
Brain
Uterine cuttering
Lymph nodes
Liver
Spleen
Soft cellular tumors
66
Cryostat mounting media under -15 to -25°C
[PNGOT]
Prostate
Non-fatty breast tissue
GI tract
Ovary
Tongue
67
Cryostat mounting media under 35°C
[FBOT]
Fatty breast
Omental tissue
68
[Cryostat]
Preferably, the tissue block
should be ___[1]___ mm. thick in order to ___[2]___
2-4
Minimize the risk of the knife hitting the metal tissue block holder
69
[cryostat]
Small fragments of tissue, such as curettings or brain biopsies, are placed on a ___[1]___
The blocks are then surrounded and covered with an ___[2]___, and frozen by ___[3]___
1. thick base of O.C.T. compound
2. additional matrix of O.C.T. compound
3. liquid nitrogen
70
The frozen tissue is mounted on the microtome. Both the microtome knife
and the tissue block are left in the cryostat for ___[1]___ minutes at ___[2]___, to ensure that they are cooled to the correct temperature
1. 15
2. -20°C
71
[cryostat]
Sections between 5-10µm are
then cut ___[1]___, removed from the knife with a ___[2]___, attached directly to slides of cover-glasses at room temperature, air dried, and fixed (optional)
1. slowly and steadily
2. camel hair brush
72
To mount cryostat sections after cutting, one edge of the glass slide is
___[1]___
The section will ___[2]___
1. lowered gently until it is about 1/2 to 1 mm. from the knife face
2. automatically transfer from the cold knife to the relatively warm slide
73
[cryostat]
The slide should never be pressed down on the section, because this will cause a
___[1]___.
If this happens, ___[2]___
1. frost mark to remain where the section rested on the knife
2. the frost mark should be wiped away with soft tissue paper
74
Overall, cryostat sections provide the simplest, quickest and least labor intensive method for producing frozen sections, and are routinely used for ______ of surgical specimen
intraoperative and rapid diagnosis
75
It should be noted that cryostats cut only individual sections, and do not
______
form ribbons
76
Cryostat sections of fresh, unfixed tissue usually ___[1]___, and will preserve ___[2]___ that may be studied by histochemical techniques
1. attach easily to the slide, even without adhesives
2. enzymes and other substances
77
The cryostat is also recommended for any technique requiring cold
sectioning of fixed material, e.g., for ___[1]___, and for some special
methods for the ___[2]___
1. fats and lipids
2. nervous system
78
[Freezing Previously Fixed Tissue]
Sections of formalin-fixed tissue, however, may not adhere to the slide, and ______
will fall off or be detached during staining
79
[Freezing Previously Fixed Tissue]
Clean slides should be coated with ___[1]___ or ___[2]___ so that the fixed tissue will attach to the slide
1. albumin
2. chrome-glycerin jelly
80
Special fixatives such as ______ may be used in histochemistry and for lipid
demonstration
10% formol calcium at 4°C
81
[Freezing Previously Fixed Tissue]
Special fixatives such as 10% formol calcium at 4°C may be used in ______
histochemistry and for lipid
demonstration
82
Tissues that have been fixed or stored in alcohol should be ___[1]___ before sectioning, since ___[2]___
1. washed in water for 12-24 hours
2. alcohol inhibits freezing
83
[Freezing Previously Fixed Tissue]
Cumbersome way to attach fixed tissue to slide is to ______ before freezing and sectioning for rapid surgical diagnosis
immerse the tissue block in boiling 10% buffered formalin for 1 to 2 minutes
84
[nerve and muscle]
The portion of a specimen intended for frozen section should be transported on ___[1]___, and rapidly frozen within ___[2]___
1. top of wet ice, on saline-dampened gauze
2. two hours
85
Muscle and nerve biopsies are divided into separate portions that will allow for formalin fixation and paraffin embedding
unfixed snap-frozen for ___[1]___
fixation and resin embedding for ___[2]___ and in some rare cases for ___[3]___
1. cryostat sections
2. electron microscopy (EM)
3. biochemical immunoblotting studies
86
[Examination of nerve and muscle]
Upon receipt in the histology lab, specimen is oriented in ___[1]___ compound and snap-frozen in liquid ___[2]___ for optimal results
1. O.C.T. (Optimal Cutting Temperature)
2. nitrogen/ isopentane
87
[Examination of nerve and muscle]
Are critical to obtaining undamaged sections of unfixed muscle fibers
Orientation, size, and expedient flash freezing
88
[Examination of nerve and muscle]
Do not allow the tissue to freeze slowly or to soak up excess saline, as these will cause:
artifacts that can be seen microscopically and can interfere with diagnostic interpretation
89
[Examination of nerve and muscle]
A portion of the tissue is oriented on a ___[1]___, fixed with ___[2]___ and processed for staining with ___[3]___
1. piece of cardboard
2. 10% buffered formalin
3. routine Hematoxylin and Eosin (H&E staining)
90
[Examination of nerve and muscle]
The biopsy portion for electron microscopy is fixed in a ___[1]___ and postfixed in ___[2]___, usually by a ___[3]___
1. buffered solution of glutaraldehyde
2. osmium tetroxide
3. specialist in electron microscopy
91
[Examination of nerve and muscle]
In some ______, biochemical techniques may also be required
muscular degenerative disorders
92
For histochemical evaluation involving enzyme studies, the tissue needs to
be ______, and the important chemical constituents should not have
been removed, altered or displaced
chemically active
93
Deemed to be the most ideal and preferred means of preserving tissues in order to avoid
complete or partial loss of enzymes consequent to chemical fixation
Frozen section
94
[SPECIAL PROCESSING TECHNIQUES]
Difficulties, however, arise in _______; since cut sections of tissue tend to disintegrate and cannot be easily handled without prior fixation
obtaining thin and serial sections of uniform thickness
95
In addition to fresh frozen tissue sectioning, there are methods that may be resorted to, if chemical fixation of tissue blocks is to be avoided, namely:
1. Freeze-drying
2. Freeze substitution
96
Like fresh frozen sections, special techniques have the common principle of rapidly preserving the tissue block by ______
freezing (quenching)
97
The aim of rapidly preserving tissue block by freezing is to ______ thereby preventing
chemical alteration of tissue and displacement of cellular tissue components
produce instant cessation of cellular activity
98
The aim of rapidly preserving tissue block by freezing is to produce instant cessation of cellular activity thereby ______
preventing chemical alteration of tissue and displacement of cellular tissue components
99
The use of isopentane, pentane and propane and most recently of dichloro-difluoromethane, which can be cooled to very low temperature in order to ______
retain the fluidity of the freezing agents
100
The use of ___[1-4]___, which can be cooled to very low temperature in order to retain the fluidity of the freezing agents
Isopentane,
Pentane
Propane
Dichloro-difluoromethane
101
A special way of preserving tissues by rapid freezing (quenching) of fresh tissue at -160°C and subsequently removing ice water molecules (dessication) by transferring the still frozen tissue block into a vacuum chamber at a higher temperature, e.g. -40°C (sublimation) without the use of any chemical fixative
Freeze-drying
102
Temp ng rapid freezing sa Freeze-Drying
-160degC
103
Temp ng vacuum chamber sa Freeze-Drying
-40degC
104
[freeze-drying]
A tissue around ___[1]___ thick is plunged into ___[2]___ mixture which has been chilled to -160° to -180°C with liquid nitrogen
1. 2 mm.
2. isopentane or propaneisopentane
105
[Freeze-Drying]
Tissues plunged into isopentane or propaneisopentane mixture which has been chilled to -160° to -180°C with liquid nitrogen will effectively solidify the tissue in _______
2-3 seconds
106
[Freeze-Drying]
Tissues plunged into isopentane or propaneisopentane mixture which has been chilled to -160° to -180°C with liquid nitrogen will effectively solidify the tissue in 2-3 seconds preventing ______
the formation of large ice crystals, autolysis and putrefaction
107
[Freeze-Drying]
Water is sublimated and dehydrated from the tissue, thereby completing the dessication process within _______.
24-48 hours
108
[Freeze-Drying]
Once drying is completed, the tissue is removed, fixed and embedded, either in ___[1]___, ___[2]___ or ___[3]___
1. molten paraffin wax
2. water soluble waxes
3. celloidin
109
[Freeze-Drying]
Infiltration and impregnation are usually performed in a ______.
vacuum embedding oven
110
By far the most time consuming part of the Freeze-Drying process
Drying
111
Why drying is the most time-consuming procedure of freeze-drying?
certain tissues contain 70- 80% water by weight that has to be removed without damage to the tissue
112
Bakit mas mahirap i-section ang freeze-dried sections compared sa paraffin blocks?
The tissue is brittle and inadequately supported due to the relatively short period for wax impregnation
113
[Freeze-Drying]
The tissues are usually flattened directly into an ______ with the aid of the finger
albuminous glass slide
114
[Freeze-Drying]
___[1]___ must be avoided and ___[2]___ are preferred
1. Water
2. warm alcohol, acetone, mercury
115
Freeze-Drying produces ___[1]___tissue shrinkage, and allows tissues to be processed in a ___[2]___ state
1. minimum
2. fresh
116
Freeze-Drying causes minimal chemical change on the cells, most especially on the ___[1]___ components, and less ___[2]___
1. protein
2. displacement of tissue and cellular constituents
117
Freeze-drying may be used for special studies, including:
1. Immunocytochemistry 2. Fluorescent antibody studies of polypeptide and polypeptide hormones
3. Autoradiography
4. Microspectrofluorimetry of autofluorescent substances
5. Formaldehyde-induced fluorescence of biogenic amines (to demonstrate 5-hydroxytryptamine, adrenaline, and other catecholamines)
6. Scanning electron microscopy
118
Freeze-drying demonstrates:
hydrolytic enzymes
mucous substances
glycogen
proteins
119
A process of dehydration, performed at temperatures low enough to avoid the formation of ice crystals and to circumvent the damaging effects observed after ambient-temperature dehydration
Freeze-substitution
120
Difference of FREEZE-DRYING and FREEZE-SUBSTITUTION
FREEZE-DRYING
Dehydrated in a vacuum
FREEZE-SUBSTITUTION
Fixed in Rossman's formula or in 1% Acetone
Dehydrated in absolute alcohol
121
Infiltration and embedding of Freeze-Substitution is then carried out in the same way as in ______
paraffin section
122
Freeze-substitution is based on rapid freezing of tissues followed by:
solution ("substitution") of ice at temperatures well below 0°C
123
Freeze-substitution
A ___[size]___ specimen is thrown into ___[2]___ that is super cooled by liquid nitrogen to ___[3]___°C (with precautions)
size. 1-3 mm
2. 3:1 propane - isopentane
3. -175
124
[freeze-substitution]
Cryostat sections are cut ___[1]___ µm, and transferred to ___[2]___ (substituting fluid), and cooled to ___[3]___degC for ___[4]___ in order to dissolve ice slowly without distorting tissue structure
1. 8-10
2. water-free acetone
3. -70
4. 12 hours to 1 week
125
This technique is relatively more economical and less time-consuming than freeze-drying
Freeze-Substitution
126
For best morphological and histochemical preservation, substituting fluids should in general contain both ______
chemical fixing agent and solvent for ice
127
Freeze-Substitution substituting fluids:
1% osmium tetroxide in acetone
mercuric chloride in ethanol
picric acid in ethanol
128
Other name of infiltration
Impregnation
129
Other name of sectioning
Cutting / Microtomy
130
Describes the various steps required to take the tissue from fixation to the state where it is completely infiltrated with a suitable histological wax and can be embedded ready for section cutting on the microtome
Tissue processing
131
______ cannot be directly infiltrated with paraffin
Wet fixed tissues (in aqueous solutions)
132
Dehydration is usually done with increasing concentrations of alcohol _____
(70% to 95% to 100%)
133
Most common clearing agent
Xylene
134
Formalin, usually as a ______ solution, is the most popular fixative for preserving tissues that will be processed for paraffin embedding
phosphate-buffered
135
Specimens should remain in fixative long enough for it to ___[1]___ and then for an additional period in order to ___[2]___
1. penetrate the tissue
2. allow the chemical reactions of fixation to reach equilibrium (fixation time)
136
If fixation is not carried out under optimal conditions or if fixation is delayed, a tissue specimen can be:
irreversibly damaged
137
Formalin-fixed, paraffin-embedded tissues may be stored ______ at room temperature
indefinitely
138
Nucleic acids (both DNA and RNA) may be recovered from them ______ after fixation.
decades
139
Why is paraffin not miscible with water
Kasi hydrophobic
140
______ are removed at lower concentrations of ethanol [dehydration]
Water soluble proteins
141
When ethanol concentration is increased to 100% [dehydration], ______ may be dissolved
certain lipids
142
A typical dehydration sequence for specimens not more than 4mm thick would be:
70% ethanol 15 min
90% ethanol 15 min
100% ethanol 15 min
100% ethanol 15 min
100% ethanol 30 min
100% ethanol 45 min
143
Ethanol is a poor ___ solvent
fat
144
To ensure complete dehydration,
a superior fat solvent such as ___[1]___ should be added before the final absolute ethanol,
and ___[2]___ used as the transition solvent
1. acetone or isopropanol
2. chloroform or trichloroethane
145
[clearing]
The dehydrated tissue is transferred to an intermediate solvent that is fully miscible with ______
both ethanol and paraffin wax
146
The term “clearing” has been chosen because many (but not all) clearing agents impart an optical clarity or transparency to the tissue due to their ______
relatively high refractive index
147
Another important role of the clearing agent is to remove ______ from the tissue which otherwise presents a barrier to wax infiltration
a substantial amount of fat
148
A typical clearing sequence for specimens not more than 4mm thick would be:
Xylene 20 min
Xylene 20 min
Xylene 45 min
149
Paraffin wax is liquid at what temp
60degC
150
Paraffin wax is allowed to cool to ___°C in order to solidify into a consistency that allows sections to be cut
20
151
Paraffin wax are mixtures of purified paraffin wax and various additives that may include resins such as ______
styrene or polyethylene
152
A typical paraffin infiltration sequence of paraffin for specimens not more than 4mm thick would be:
Paraffin wax 30 min
Paraffin wax 30 min
Paraffin wax 45 min
153
The infiltrated tissue is removed from the cassette and very carefully oriented in a suitably sized metal mold so that the “______” can be determined
plane of section
154
Usually tissues are embedded with the surface to be cut facing ______
down in the mold
155
The choice of mold will depend on the ______ that will be used to section the tissue.
type of chuck in the microtome
156
The process by which tissues are first embedded or fully infiltrated with a supporting medium such as agar or nitrocellulose, then infiltrated a second time with wax in which they are also embedded
Double embedding
157
Double embedding is the process by which tissues are first embedded or fully infiltrated with a supporting medium such as ______, then infiltrated a second time with wax in which they are also embedded
agar or nitrocellulose
158
Double embedding in agar-paraffin is a reliable and convenient method of handling minute and friable tissue fragments such as ______
curetting and endoscopic biopsies
159
The tissues may ______ by the time they are infiltrated with wax, but if adequately processed, they will still show ______ l and allow for accurate histopathological evaluation
1. shrink
2. good morphological detail
160
Specimens for routine Hematoxylin and Eosin (H&E) are cut _____ μm in thickness
3–5
161
Tissues to be examined for amyloid
deposits are better sectioned at _____ μm
8–12
162
Kidney biopsies should be cut
at _____ μm for optimal viewing of the structures of glomeruli
2
163
The most commonly employed embedding media for semi-thin and ultrathin sections
Epoxy resins
164
Acrylic resins are also used as embedding media, particularly where ______ is required
immunohistochemistry
165
Thicker sections (0.35μm to 5μm) of resin-embedded tissue can also be cut for ______
light microscopy
166
The immiscibility of most epoxy and acrylic resins with water necessitates the use of ______ , usually with ethanol.
dehydration
167
Knives are either of the standard ______ variety or ______ variety
thick metal
thin disposable
168
Usually this distance can be set, for most paraffin embedded tissues at ______ microns
6 to 8
169
Plastic blocks (methacrylate, araldite, or epon) are sectioned with ___ or _____ that can cut sections down to about ___ micron
glass
diamond knives
1
170
Example of plastic blocks
(methacrylate, araldite, or epon)
171
Thin sections for electron microscopy (0.25 micron) are best done with a ______ knife
diamond
172
The glass slides containing the specimen are then placed in a warm oven [after sectioning] for about ___ minutes to help the section adhere to the slide
15
173
If heat from oven (after microtomy) might harm such things as antigens for immunostaining, then this step can be bypassed and ______ can be used instead to pick up the sections
glue-coated slides
174
The temperature of the warm bath should be kept at ___ degC below the melting point of the embedding wax
5–10
175
If waterbath is too hot, ______ looking sections will result, while cool water baths will produce ______ of the tissue
desiccated-
excessive wrinkling
176
Adding a few drops of ______ to the water [mounting] may facilitate the mounting of tissue sections
ethyl alcohol
177
The ribbon must not be left in the bath for more than ______ minutes, or ______ of the tissue will be produced
1 or 2
over-hydration
178
Because tissue sections do not adhere well to untreated glass slides, a ______ also must be a component of the water bath. ______, and ______ are all suitable additives of this type
bonding agent
Albumin, poly-L-lysine
179
Tissue adheres to a wooden applicator stick that is used to float successively prepared ribbons from two different cases
tongue blade metastasis
180
In cases where the specimen is limited in size (such as ______, ______, ______), it is advisable to save any unmounted paraffin ribbon (with appropriate identification) from such cases for at least _____ after they are accessioned
1. skin, bronchoscopy, gastrointestinal biopsies
2. week
181
The tissue is ______ during the first steps in preparing the slides for staining
deparaffinized
182
Because the stainer baths are a potential reservoir of tissue contaminants, changing the staining fluids may alleviate some of the potential for ______ from this source
carryover
183
If the processor is to be run overnight, it should be programmed to hold on the ______ and not finish until the next morning so the specimens do not sit in hot paraffin longer than the time indicated
first ethanol bath
184
If specimens are fresh they may incubate in ______ in the first stage on the (automatic tissue processing) machine
formalin
185
It is important to not keep the tissues in hot paraffin too long or else they may become ______
hard and brittle
186
Processed tissues can be stored in the cassettes at room temperature ___[time period]___
indefinitely
187
Is the impregnation of tissues by a molten medium under reduced pressure
Vacuum infiltration
188
The procedure assists the complete and rapid impregnation of tissues with wax and reduces the time tissues are subjected to high temperatures, thereby minimizing heat-induced tissue hardening, facilitating complete removal of transition solvents, and prolonging the life of wax by reducing solvent contamination
Vacuum infiltration
189
Vacuum infiltration assists the complete and rapid impregnation of tissues with wax and reduces the time tissues are subjected to high temperatures, thereby:
minimizing heat-induced tissue hardening,
facilitating complete removal of transition solvents,
prolonging the life of wax by reducing solvent contamination
190
Factors that impact the duration of tissue processing and extent of infiltration
1. Tissue Density and Thickness
2. Agitation
3. Temperature
4. Vacuum and pressure
191
Agitation using manual or automated processors increases the ______ in and around the tissues
flow of fresh fluids
192
Most tissue processing protocols utilize automated processors with ______ oscillation mechanisms to speed fluid exchange
vertical or rotary
193
Without agitation, tissues tend to ______ or become ______, therefore reducing surface area available for fluid exchange
settle to the bottom of the processing device
too tightly packed
194
Fluid interchange between processing reagents and tissues is promoted by ______
exposure of the maximum tissue surface area
195
Tissues should be ______ to facilitate exchange of reagents and increase diffusion
loosely packed in baskets
196
Ideally, the cassette perforations should be ______ to the fluid flow
perpendicular
197
Temperatures in the range of ______°C, for a limited time can speed up fluid penetration and tissue processing protocols
37° to 45
198
High temperature can cause the tissue to _____ and to become _____
shrink
hard and brittle
199
Low temperature increases the ______ in tissue processing, thereby reducing the rate of diffusion
viscosity of reagents used
200
Tissue shrinkage during infiltration in paraffin wax results mainly the effect of heat on ______
collagen
201
Decreased rate of diffusion in automated tissue processing can be avoided by
Maintaining embedding waxes 2-3°C above their melting points
202
Reduced pressure can ______ the infiltration rate
increase
203
High pressure facilitates infiltration of dense specimens with the more ______ embedding media
viscous
204
______ during tissue infiltration improves processing quality and can aid in removal of trapped air from porous tissue
Vacuum application
205
Using vacuum during tissue processing protocols can ______ the infiltration time when dealing with dense and fatty tissue specimens
reduce
206
Using vacuum during tissue processing protocols can reduce the infiltration time when dealing with ______ tissue specimens
dense and fatty
207
Vacuum applied during dehydration, clearing and infiltration improves the quality of processing in tissues such as ______
lung which becomes de-aerated during the process
208
duration of wax infiltration is dependent upon ______ and is not generally reduced by applying a vacuum.
viscosity
209
______ and ______ will prevent the cells from detaching during staining
Gentle washing and minimal thickness of cell layers
210
In general, _____ and _____ should be incubated conservatively
needle biopsies and bloody specimens
211
______ specimens can be processed for longer than
average
fatty
