7.3-STAINING E-F IHHH Flashcards
1
Q
What are the three protective coats covering the Central Nervous System
A
Dura mater; Arachnoid mater; Pia mater
2
Q
What are the components of the Central Nervous System
A
Brain and spinal cord
3
Q
What lines the ventricles and central canal in the Central Nervous System
A
Ependymal cells
4
Q
What are the components of Gray matter in the Central Nervous System
A
Neurons and supporting cells
5
Q
What are the components of White matter in the Central Nervous System
A
Myelinated axons and oligodendrocytes
6
Q
What is a Neuron
A
Excitable cell responsible for processing and transmitting information
7
Q
What does the Cell body of a neuron contain
A
Various subcellular organelles responsible for metabolic upkeep of cell
8
Q
What is the function of the Nucleus of a neuron
A
Storage of cell’s genetic code in DNA
9
Q
What is the function of Dendrites in neurons
A
Complex cell processes which receive synaptic inputs from other neurons
10
Q
What is the function of Axon in neurons
A
Elongated fiber which transmits electrical impulses away from the soma
11
Q
What are Nissl bodies
A
Cytoplasm
12
Q
What are the components of the cytoskeleton
A
Microtubules; Neurofilaments; Microfilaments
13
Q
What is a Neurofibril
A
A complex network of fibrils running through the cytoplasm and processes of neurons
14
Q
What are Myelin sheaths
A
Elaborated and maintained by oligodendrocytes
15
Q
What is the function of Myelin sheaths
A
Insulating layer that modifies nerve conduction along axons
16
Q
What are the Supporting elements of CNS
A
Neuroglia
17
Q
List the Neuroglia
A
Astrocytes; Oligodendrocytes; Microglia
18
Q
What is the role of Astrocytes
A
Major supporting cells in the brain
19
Q
What do Astrocytes maintain
A
Extracellular ion and neurotransmitter balance
20
Q
What is the role of Astrocytes in response to injury
A
Repair and scaring responses; respond to injury by producing a dense network of processes
21
Q
What is the role of Astrocytes in the blood-brain barrier
A
Part of blood-brain barrier
22
Q
What is the composition of glial “scar”
A
Predominantly of cytoplasmic processes; with little or no extracellular protein
23
Q
What is Astrocyte swelling often associated with
A
Increased synthesis of glial fibrillary acidic protein (GFAP); the astrocyte’s major cytoskeletal protein
24
Q
What are the characteristics of Oligodendrocytes
A
Small; rounded lymphocyte-like nuclei
25
What is the function of cytoplasmic processes of Oligodendrocytes
Wrap around the axons of neurons to form myelin
26
What are the characteristics of Oligodendrocytes compared to Astrocytes
Smaller than astrocytes and have a denser nucleus and cytoplasm
27
Where are Oligodendrocytes seen
Adjacent to myelinated nerve fibers in the white matter or forming satellite cells to the neurons in the gray matter
28
What are the characteristics of Microglia
Smallest cells of the neuroglia
29
Where are Microglia frequently seen
In gray matter in close proximity to neurons
30
What are Microglia derived from
Circulating; major phagocytic cells in the CNS
31
What is the role of Microglia
Involved in the repair of tissue damage; may accumulate abundant intracellular lipids to form foamy macrophage
32
What are Microglial nodules indicative of
Viral infections
33
List the fixatives for nervous tissue
Formol-saline; Buffered formalin; Lillie’s calcium acetate formalin (for myelin histochemistry)
34
What type of sections are used for Lipid histochemistry
Frozen sections
35
What is the recommended fixation time
At least 24 hours
36
What are Shrinkage or fixation artifacts
Perivascular “retraction” spaces
37
List the Staining techniques
Hematoxylin and Eosin; Silver impregnation methods; Basic aniline dyes (Nissl stains) – cresyl violet; gallocyanin; toluidine blue; Immunohistochemistry
38
What are Nissl bodies
Coarse granules made up of ribonucleic acid that are scattered in the cytoplasm of nerve cells
39
What does absence of Nissl bodies suggest
Nerve cell degeneration
40
What allows the stain to work on them in Nissl bodies
Rough endoplasmic reticulum has ribosomes that allow the stain to work on them
41
What are Nissl bodies best stained by
Basic dyes such as cresyl fast violet; toluidine blue; methylene blue or thionine
42
What color is Nissl substance in Cresyl fast violet (nissl) stain for paraffin sections
Purple-dark blue
43
What color are Neurons in Cresyl fast violet (nissl) stain for paraffin sections
Pale purple blue
44
What color are Cell nuclei in Cresyl fast violet (nissl) stain for paraffin sections
Purple blue
45
What are examples of pathologies shown using Cresyl fast violet (nissl) stain
Central chromatolysis; Amyotrophic Lateral Sclerosis
46
What does the staining of Astrocytes use
Tinctorial stain
47
What is the Fixation for Cajal’s gold sublimate method for astrocytes
Cajal's Formol Ammonium Bromide for 2-25 days (average 5 days)
48
What type of sections are used in Cajal’s gold sublimate method for astrocytes
Frozen Sections
49
What is the Staining Solution in Cajal’s gold sublimate method for astrocytes
Gold Sublimate
50
What color are Astrocytes in Cajal’s gold sublimate method
Bluish black on a light brownish background
51
What color are Nerve Cells in Cajal’s gold sublimate method
Red
52
What color are Nerve fibers in Cajal’s gold sublimate method
Unstained
53
What does Modified ptah stain for reactive astrocytes (neuroglial fibers) stain
Reactive astrocytes
54
What fixative is used in Modified ptah stain for reactive astrocytes (neuroglial fibers)
10% buffered or unbuffered formalin
55
What type of Sections are used in Modified ptah stain for reactive astrocytes (neuroglial fibers)
Paraffin; sectioned at 8 microns
56
What Solutions are used in Modified ptah stain for reactive astrocytes (neuroglial fibers)
Acid Zenker's solution; Phosphotungstic acid hematoxylin (PTAH)
57
What is the Fixation in Modified holzer’s method for astrocytic processes
Formalin; Helly's or Bouin's fixative
58
What type of Sections are used in Modified holzer’s method for astrocytic processes
Paraffin 6 to 10 μm
59
What Solutions are used in Modified holzer’s method for astrocytic processes
Mordant; Chloroform-Alcohol mixture; Crystal violet stain; Differentiating solution
60
What color are Glial fibrils in Modified holzer’s method for astrocytic processes
Blue
61
What color are Nuclei in Modified holzer’s method for astrocytic processes
Pale blue
62
What color is the Background in Modified holzer’s method for astrocytic processes
Colorless
63
How are Normal oligodendrocytes stained
Immuno-histochemical methods using antibodies to galactocerebroside; myelin basic protein; or carbonic anhydrase-C
64
How are Microglial cells stained
Express macrophage markers by immunocytochemistry; and will stain positively for CD68 antigens; and faintly positive with leukocyte common antigen (CD45)
65
What is the Myelin sheath
Fatty insulating sheath surrounding the axon
66
What is the Myelin sheath made up of
Lipid (cerebrosides; phospholipids; and cholesterol)
67
What are the Histochemical reactions for Myelin sheath
Baker’s chromatin-acid hematein; osmium tetroxide
68
What are the Fat stains for Myelin sheath
Sudan dyes
69
What are the Dye-metal complexes for Myelin sheath
Iron Hematoxylin; Marchi’s reaction
70
What stain is most frequently used for Myelin sheath
Luxol fast blue
71
How is Nervous tissue treated in Weigert-Pal technique of staining normal myelin sheaths
Subjected to prolonged chromation (and also to treatment with chromium fluoride and copper acetate) before dehydration and embedding in paraffin
72
How are Sections stained in Weigert-Pal technique of staining normal myelin sheaths
With an oxidized hematoxylin solution to produce blue–black deposits at the sites of the bound metal; and excess dye is removed with a borax ferricyanide solution
73
What is the duration of Weigert-Pal technique of staining normal myelin sheaths
6 weeks or more
74
What is the Fixation in Kluver & Barrera luxol fast blue with Nissl counterstain
Formalin
75
What type of Sections are used in Kluver & Barrera luxol fast blue with Nissl counterstain
Paraffin
76
What Solutions are used in Kluver & Barrera luxol fast blue with Nissl counterstain
Luxol Fast Blue (Copper phthalocyanine dye); Cresyl violet; Cresyl violet differentiator
77
What color is Myelin in Kluver & Barrera luxol fast blue with Nissl counterstain
Blue/green
78
What color is Myelin in Luxol fast blue-H&E
Blue-green
79
What color are Nuclei in Luxol fast blue-H&E
Dark blue
80
What color is Cytoplasm in Luxol fast blue-H&E
Various shades of pink
81
What color is Myelin in Luxol fast blue-PAS-Hematoxylin
Blue
82
What color are Fungi and PAS-positive elements in Luxol fast blue-PAS-Hematoxylin
Rose to red
83
What color are Nuclei in Luxol fast blue-PAS-Hematoxylin
Dark blue
84
What color are Cytoplasmic nucleoproteins in Luxol fast blue-PAS-Hematoxylin
Bluish purple
85
What color are Capillaries in Luxol fast blue-PAS-Hematoxylin
Red
86
What are mixed in the staining solution in Weil’s method
Mordant and dye
87
What are the mordant and dye in Weil’s method
Ferric ammonium sulfate (iron alum) and oxidized hematoxylin
88
What is the Fixation in Weil’s method
Formal saline or formol calcium
89
What type of Sections are used in Weil’s method
Paraffin; frozen
90
What are the Solutions used in Weil’s method
Solution A (4% Aqueous iron alum); Solution B (1% alcoholic hematoxylin); Differentiator (Weigert's borax ferricyanide)
91
What color is Myelin in Weil’s method
Black
92
What color is the Background in Weil’s method
Yellow
93
How are mounted cryostat sections treated in Baker’s chromate acid hematin
With calcium dichromate
94
What happens to Chromium in Baker’s chromate acid hematin
Bound at the sites of choline containing lipids (lecithin; sphingomyelin) and demonstrated by forming a colored complex with hematein
95
How is differentiation achieved in Baker’s chromate acid hematin
In borax-ferricyanide; removes excess hematein
96
What color are Myelin sheaths; mitochondria and erythrocytes in Baker’s chromate acid hematin
Dark blue
97
What color is Degenerating Myelin in Swank & Davenport’s marchi method for degenerating myelin
Black (early and late products)
98
What color is the Background in Swank & Davenport’s marchi method for degenerating myelin
Colorless to pale brown
99
What does Swank & Davenport’s marchi method for degenerating myelin minimize
Sporadic staining of normal myelinated fibers
100
What type of technique is Microwave Modification of Bielschowsky’s Technique for Nerve Fibers
Impregnation method; depends on reduction by formalin of silver ammonium hydroxide
101
What happens in Microwave Modification of Bielschowsky’s Technique for Nerve Fibers
Stains axons and neuronal processes by using a primary "sensitizing" solution; followed by impregnation in ammoniacal silver solution to intensify the result; and final reduction in formalin
102
What does Microwave Modification of Bielschowsky’s Technique for Nerve Fibers consist of
A silver nitrate combination that is toned with gold chloride to demonstrate nerve fiber
103
What color are Cytoplasmic neurofibrils in Microwave Modification of Bielschowsky’s Technique for Nerve Fibers; Neurofibrillary Tangles and Senile Plaques
Brown to black
104
What color are Neurofibrillary tangles and Plaques in Microwave Modification of Bielschowsky’s Technique for Nerve Fibers; Neurofibrillary Tangles and Senile Plaques
Dark brown or black
105
What color is Neuromelanin in Microwave Modification of Bielschowsky’s Technique for Nerve Fibers; Neurofibrillary Tangles and Senile Plaques
Black
106
What color is Lipofuscin in Microwave Modification of Bielschowsky’s Technique for Nerve Fibers; Neurofibrillary Tangles and Senile Plaques
Brown or black
107
What is used in Bodian stain for nerve fibers and nerve endings
Protargol-S (silver proteinate) is used with the addition of copper metal
108
What does Copper do in Bodian stain for nerve fibers and nerve endings
Replaces silver in connective tissue; allows greater differentiation between nerve fibers and connective tissue
109
How are Sections treated in Bodian stain for nerve fibers and nerve endings
Toned with gold chloride
110
What does Bodian stain for nerve fibers and nerve endings demonstrate
Neuritic plaques and neurofibrillary tangles; for the diagnosis of Alzheimer's disease
111
What does Sevier-Munger Technique for Staining of Neural Tissues stain
Neuritic plaques and neurofibrillary tangles in brain of patients with Alzheimer's disease
112
What is the procedure in Sevier-Munger Technique for Staining of Neural Tissues
Similar to Bielschowsky technique; except that the tissue is impregnated with an ammoniacal silver solution instead of a silver nitrate combination
113
What color are Large and small peripheral neurites in Sevier-Munger Technique for Staining of Neural Tissues
Black
114
What color are Axons in Sevier-Munger Technique for Staining of Neural Tissues
Black
115
What color is Myelin sheath in Sevier-Munger Technique for Staining of Neural Tissues
Light brown
116
What color are Neuritic plaques and tangles in Sevier-Munger Technique for Staining of Neural Tissues
Black
117
What color are Argentaffin granules in Sevier-Munger Technique for Staining of Neural Tissues
Black
118
What is demonstrated in Golgi’s silver staining technique
By impregnating fixed nervous tissue with potassium dichromate and silver nitrate
119
What is the result in Golgi’s silver staining technique
Black deposit on surface of nerve cells and neuroglia in colorless background
120
What is done in Modified golgi method
Addition of vacuum
121
What is the purpose of Addition of vacuum in Modified golgi method
Removal of air from tissue permits better penetration of reagents
122
What is Glial fibrillary acidic protein (GFAP)
An intermediate filament protein that is expressed by numerous cell types of the CNS including astrocytes and ependymal cells
123
What does GFAP act as
Cell-specific marker; distinguishes astrocytes from other glial cells
124
What does Upregulation of GFAP expression indicate
Astrocytic reaction commonly observed after CNS lesion
125
Where can GFAP be demonstrated
In Schwann cells; enteric glial cells and satellite cells of human sensory ganglia
126
How is GFAP demonstrated in neoplastic tissues
This antibody is useful for the identification of astrocytomas and ependymomas
127
How is GFAP stained
By immunohistochemical methods and immunofluorescence
128
What is the Staining Pattern of GFAP ANTIBODY STAINING FOR IHC
Cytoplasmic (glial cells such as astrocytes and ependymal cells)
129
What does NeuN (or Neuronal Nuclei) react with
Most neuronal cell types throughout the central and peripheral nervous systems
130
What is NeuN (or Neuronal Nuclei) an excellent marker for
Neurons in primary cultures and in retinoic acid-stimulated P19 cells as well as for identifying neurons in transplants
131
What is the Staining Pattern of NEUN ANTIBODY STAINING FOR IHC
Nuclear/cytoplasmic (primarily in nuclei with lighter staining in the cytoplasm)
132
What is Myelin basic protein (MBP)
Major constituent of the myelin sheath of oligodendrocytes and Schwann cells in the central nervous system and the peripheral nervous system; respectively
133
What is the Staining Pattern of MBP ANTIBODY STAINING FOR IHC
Fibrillar staining pattern – normal brain
134
What does the PERIPHERAL NERVOUS SYSTEM consist of
Group of neurons (ganglion cells) and network of nerve fibers (plexuses); nerves; nerve roots
135
What are the divisions of the PERIPHERAL NERVOUS SYSTEM
Somatic; Visceral
136
What fixative is used for electron microscopy of peripheral nerve segments
2.5–3% glutaraldehyde
137
What fixative is used for immunohistochemistry of peripheral nerve segments
Paraformaldehyde is generally used in concentrations of 1–4%
138
How long should Peripheral nerve segments be immersed in the fixative
For 10–12 hours
139
Where should samples immersed in glutaraldehyde or paraformaldehyde be kept
In the refrigerator
140
How are samples post-fixed
In 1% osmium tetroxide for two hours at room temperature; embedded in epoxy resin in cross and longitudinal orientation; and then sectioned at 1-μm thickness and stained with methylene blue
141
What do myelin sheaths appear as in H&E stained Peripheral myelin in paraffin sections
Largely empty zones surrounding eosinophilic axons and encircled by eosinophilic Schwann cell cytoplasm and other components of the endoneurium
142
What techniques can be used to visualize Peripheral myelin in paraffin sections
Dye metal complexes of iron; Water insoluble anionic dyes
143
What color is Myelin in Methylene blue-azure ii-basic fuchsin
Blue
144
What color are Other tissue elements in Methylene blue-azure ii-basic fuchsin
Light blue
145
What color is Collagen in Methylene blue-azure ii-basic fuchsin
Pink/red
146
What color is Elastin in Methylene blue-azure ii-basic fuchsin
Red
147
What does Osmium tetroxide provide
Insoluble; visible deposits at sites of unsaturation
148
When is Osmium tetroxide used
As a primary fixative or for post-fixation after the addition of formaldehyde or glutaraldehyde
149
What is Osmium tetroxide
Strong oxidizing agent; OsO4 reacts with a wide range of organic compounds and is itself reduced to a black lower oxide; OsO2
150
What are the basic components of proteins,carbon
hydrogen
151
What are simple proteins based on chemical composition,albumins
globulins
152
What are conjugated proteins based on chemical composition,lipoproteins
mucoproteins
153
What are derived proteins based on chemical composition,
denatured proteins and peptides
154
What are fibrous proteins based on physical configuration,actin
collagen
155
What is a common feature of fibrous proteins,Often structural
provides strength to tissues
156
Give three examples of staining methods for fibrous proteins,Trichrome method
Reticulin method
157
What are globular proteins based on physical configuration,Water soluble
many are enzymes
158
What are the functions of globular proteins,transporting
catalyzing
159
What are some examples of globular proteins,albumins
alpha globulin
160
What are membrane proteins based on physical configuration,relaying signals within cells
allowing cells to interact
161
What is another function of membrane proteins,
serve as receptors or provide channels for polar or charged molecules to pass through the cell membrane
162
Give examples of membrane proteins,c-myc
estrogen receptor
163
What do nucleic acids usually combine with,
basic proteins to form nucleoproteins
164
What do nucleic acids consist of,alternate sugar and phosphate groups
with a nitrogenous base attached to each sugar group
165
What is the 5-carbon sugar in DNA,
deoxyribose
166
Where is DNA mainly found,
in the nucleus of the cell
167
What are the four nitrogenous bases of DNA,
purines (adenine and guanine) and pyrimidines (thymine and cytosine)
168
What is the 5-carbon sugar in RNA,
ribose sugar
169
Where is RNA found,
in the cytoplasm and to a lesser extent in the nucleus
170
What are the nitrogenous bases of RNA,
purines (adenine and guanine) and pyrimidines (uracil and cytosine)
171
What are acidic dyes (anionic/negatively charged) stain,acidophilic substances which are cationic (positively charged) at physiologic pH
172
What are basic dyes (cationic/positively charged) stain,basophilic substances
which are anionic (negatively charged) at physiologic pH
173
What does Hematoxylin and eosin involve,application of hemalum
a complex formed from aluminum ions and hematin (an oxidation product of hematoxylin)
174
What is nuclear staining followed by in Hematoxylin and eosin,counterstaining with an aqueous or alcoholic solution of eosin Y
which colors eosinophilic structures in various shades of red
175
What are histochemical identification of proteins based upon,
identification of specific linkages or groups within the amino acid molecule
176
What are some specific linkages or groups within the amino acid molecule used for histochemical identification of proteins,
protein bound amino groups (e.g. in lysine) phenyl groups (e.g. in tyrosine)
177
What fixative is most commonly used for amino acid histochemistry,
Neutral buffered formol saline
178
What embedding medium is commonly used for amino acid histochemistry,
Plastic embedding medium (glycol methacrylate)
179
What staining is used for ribboned epon sections 0.3-2 μ thick for light microscopy,two intense acid dyes
Aniline Blue Black and Coomassie Brilliant Blue R 250
180
What type of dye is used in Alkaline Fast-Green Method for Basic Proteins,
acid dye
181
What does Alkaline Fast-Green Method for Basic Proteins stain,
basic groups in the tissues particularly basic protamines and histones which have higher isoelectrical points than the pH of the staining solution
182
What does Peracetic Acid oxidize in Peracetic Acid-Alcian Blue for Cystine and Cysteine,
cystine and cysteine forming strong cysteic acid
183
What stains the strong cysteic acid formed by Peracetic Acid-Alcian Blue for Cystine and Cysteine,
a basic dye stains the strong cysteic acid blue-green
184
What is stained first in Alcian Blue-PAS Staining for Proteoglycans,
staining all the acidic mucins with Alcian blue
185
What happens to those acidic mucins which are also PAS positive in Alcian Blue-PAS Staining for Proteoglycans,
will be chemically blocked and will not react further during the technique
186
What is demonstrated in a contrasting manner in Alcian Blue-PAS Staining for Proteoglycans,
neutral mucins which are solely PAS positive will subsequently be demonstrated
187
What does Feulgen technique demonstrate in Staining of nucleic acids,
sugar
188
What does Methyl green pyronin technique demonstrate in Staining of nucleic acids,
phosphate
189
What demonstrates both DNA and RNA in Staining of nucleic acids,
Gallocyanin-chrome alum method
190
What is removed from the DNA by Acid hydrolysis in Feulgen Staining for Nuclear DNa,
purine bases thereby unmasking free aldehyde groups
191
What acid is used in Feulgen Staining for Nuclear DNa,
1M HCl at 60oC hydrolyze and break purine-deoxyribose bond
192
What does Feulgen reaction allow in Feulgen Staining for Nuclear DNa,
DNA in situ to be specifically stained based on the reaction of Schiff or Schiff-like reagents with the free aldehyde groups in proportion to the DNA concentration in the cell which results in the purple staining
193
Why is the Feulgen reaction DNA-specific,
RNA is not hydrolyzed by the HCl treatment
194
List some acceptable fixatives for Feulgen Staining for Nuclear DNa,
Zenker’s Carnoy’s
195
What color is DNA in Methyl Green-Pyronin method for RNA and DNA,
Green or blue-green
196
What color is Cytoplasm in Methyl Green-Pyronin method for RNA and DNA,
green
197
What color is RNA in Methyl Green-Pyronin method for RNA and DNA,
Rose-red Granules Dark rose-red
198
What fixatives are acceptable in Methyl Green-Pyronin method for RNA and DNA,
10% Formalin absolute alcohol
199
What is the most widely used fluorochrome for Fluorescent Staining for DNA and RNA,
Fluorescein
200
Why is Fluorescein the most widely used fluorochrome for Fluorescent Staining for DNA and RNA
,because of its wide absorption spectrum and blue light range
201
What type of emission is characteristic of Fluorescein,
Characteristic apple green emission is rarely seen as "auto fluorescence" in mammalian tissue which is often blue in color
202
What light does Rhodamine conjugates absorb maximally in Fluorescent Staining for DNA and RNA,
in green light
203
What emission does Rhodamine conjugates exhibit in Fluorescent Staining for DNA and RNA,
an orange-red emission
204
When are Rhodamine conjugates commonly used in Fluorescent Staining for DNA and RNA,
in two-color techniques
205
What type of light does Acridine Orange absorb in Fluorescent Staining for DNA and RNA,
Most commonly used fluorochrome to demonstrate DNA and RNA in fresh or fixed tissues combining with nucleic acids in cells by salt linkages and cohesion
206
What color does DNA emit in Fluorescent Staining for DNA and RNA using Acridine Orange,
a yellow-green fluorescence
207
What color is RNA stained in Fluorescent Staining for DNA and RNA using Acridine Orange
Brilliant orange; red; or brick to orange-red
208
What is the use of Acridine Orange in Fluorescent Staining for DNA and RNA
Screening of cervical smears for cancer cells
209
What is Acriflavine used as in Fluorescent Staining for DNA and RNA
As a 0.01% alcoholic solution; alternative to basic fuchsin in Schiff's reagent; for the Feulgen technique of acid hydrolysis
210
What color is DNA stained in Fluorescent Staining for DNA and RNA using Acriflavine
Fluorescent yellow color in this Feulgen-type reaction
211
What does Phosphotungstic acid in aqueous solution serve as in Electron microscopy
Single and reliable stain for aldehyde-fixed tissue
212
What is Polyacrylamide Gel Electrophoresis
Technique widely used to separate biological macromolecules; usually proteins or nucleic acids; according to their electrophoretic mobility
213
What may the gel be stained with after Polyacrylamide Gel Electrophoresis
For proteins; most commonly with Coomassie Brilliant Blue R-250; ethidium bromide; or silver stain
214
What is In-situ hybridization
Most sensitive technique for identifying DNA
215
What does In-situ hybridization use
A labeled complementary DNA; RNA or modified nucleic acids strand (or probe) to localize a specific DNA; RNA; nucleic acid sequence or gene expression within a cell
216
What does In-situ hybridization involve
Pre-treatment of cellular preparations to unmask the target nucleic acids; and hybridization of a nucleic acid probe of complementary base sequence to the target cells
217
How can the results of In-situ hybridization be visualized
Fluorescence (FISH) and by chromogenic (CISH) detection
218
What is the targeted treatment for Her2 neu
Herceptin (tratuzumab)
219
What can Polymerase chain reaction do
Single or few copies of a piece of DNA can be amplified generating thousands to millions of copies
220
What does Polymerase chain reaction rely on
Thermal cycling
221
What are primers
Short DNA fragments containing sequences complementary to the target region along with DNA polymerase
222
What are the three major steps of Polymerase chain reaction
Denaturation; Primer annealing; Extension
223
What happens during Denaturation in Polymerase chain reaction
95oC; break weak hydrogen bonds; creating separate single stranded DNA
224
What happens during Primer annealing in Polymerase chain reaction
54oC; primers bind (anneal) to their complementary single-stranded DNA template; polymerase attaches and starts copying the template
225
What happens during Extension in Polymerase chain reaction
72oC; optimal temperature for DNA polymerase to act; DNA building blocks complementary to the template are coupled to the primer; making a double stranded DNA molecule and adding nucleotides onto the primer in sequential manner
226
What is Reverse transcription polymerase chain reaction (rt-pcr)
Most sensitive technique for mRNA detection and quantitation
227
What happens to RNA template in Reverse transcription polymerase chain reaction (rt-pcr)
RNA template is first converted into a complementary DNA using reverse transcriptase
228
What does Reverse transcription polymerase chain reaction (rt-pcr) use
Reverse transcriptase and primer to anneal and extend a desired mRNA sequence
229
What does Reverse transcriptase and primer do to the mRNA sequence in Reverse transcription polymerase chain reaction (rt-pcr)
Reverse transcriptase and primer will anneal to the mRNA sequence and transcribe a complementary strand of DNA
230
What is Strand is replicated with in Reverse transcription polymerase chain reaction (rt-pcr)
Strand is replicated with primers and Taq polymerase; and the standard PCR protocol is followed
231
What is In-situ pcr
Powerful method that detects minute quantities of rare or single-copy number nucleic acid sequences in frozen or paraffin-embedded cells or tissue sections for the localization of those sequences within the cells
232
What takes place in the cell on a slide in In-situ pcr
Polymerase chain reaction actually takes place in the cell on a slide; and the product can be visualized in the same way as in traditional in situ hybridization
233
What can be detected through the combined examination of PCR/RT-PCR and Histology in In-situ pcr
Either DNA or RNA can be detected
