F3-IMMUNOHISTOCHEMISTRY AND IMMUNOFLUORESCENCE Flashcards
(316 cards)
1
Q
Which technique uses the principles of immunology by detecting antigens and antibodies
A
Immunohistochemistry + Immunofluorescence
2
Q
Which technique uses enzymes and chromogens as labels to produce colored products visible by light microscope
A
Immunohistochemistry
3
Q
Which technique uses fluorochromes as labels to emit light and requires fluorescence microscopy
A
Immunofluorescence
4
Q
Which technique identifies cellular or tissue constituents by antigen-antibody interactions
A
Immunohistochemistry
5
Q
Which methods are used to determine antigens in immunohistochemistry
A
Direct labeling + Secondary labeling indirect
6
Q
Which use of immunohistochemistry involves highlighting the presence of antigens in tissue
A
Diagnosis
7
Q
Which use of immunohistochemistry helps differentiate tumors with similar morphology
A
Diagnosis
8
Q
Which use of immunohistochemistry evaluates proliferation potential
A
Evaluation of proliferation potential
9
Q
Which use of immunohistochemistry identifies infectious agents in tissue sections
A
Identification of infectious agents
10
Q
Which use of immunohistochemistry provides information about patient outcome and treatment
A
Prognostic and therapeutic implications
11
Q
Which prognostic indicator is used in breast cancer
A
HER2Neu
12
Q
Which breast cancer patients have a higher chance of longer lifespan or cure
A
ERPR double positive
13
Q
Which breast cancer patients have a poorer prognosis
A
ERPR triple negative
14
Q
Which therapy is used for ERPR positive breast cancer patients
A
Hormonal therapy
15
Q
Which drugs are used for HER2Neu positive breast cancer patients
A
Herceptin + Trastuzumab
16
Q
Which use of immunohistochemistry is important for drug development
A
Drug development
17
Q
Which use of immunohistochemistry is important for biological research
A
Biological research
18
Q
What is an antigen
A
Molecule that induces antibody formation and bears antibody-binding sites
19
Q
What is an epitope
A
Highly specific topographical region composed of a small number of amino acids or monosaccharide units
20
Q
What is an antibody
A
Class of serum proteins known as immunoglobulins
21
Q
What is the basic unit of an antibody
A
Monomer
22
Q
Which immunoglobulin is most frequently used in immunohistochemistry
A
IgG
23
Q
What principle describes the binding of antibody to antigen
A
Lock and key principle
24
Q
What is the specific region on an antigen that binds to the antibody
A
Epitope
25
What is the specific region on an antibody that binds to the antigen
Paratope
26
What is affinity
Three-dimensional fit of the antibody to its specific antigen
27
What does affinity measure
Binding strength between antigenic epitope and antibody-combining site
28
What is avidity
Functional combining strength of an antibody with its antigen due to heterogeneity of antiserum
29
What does avidity describe
Heterogeneity of antiserum containing various antibodies reacting with different epitopes
30
What is antibody specificity
Characteristic of antibody to bind selectively to a single epitope on an antigen
31
What is sensitivity in immunohistochemistry
Relative amount of antigen that the technique can detect
32
Which color is most common in immunohistochemistry
Brown
33
Which chromogen is most common and produces brown color
DAB
34
Which technique allows visualization of antigen distribution in tissue or cells
Immunohistochemistry + Immunofluorescence
35
Which microscopy is used for immunofluorescence
Fluorescence microscope
36
Which microscopy is used for immunohistochemistry
Light microscope
37
Which labeling method uses a single fluorescent antibody
Direct immunofluorescence
38
Which labeling method uses an unlabeled primary antibody and a fluorescent secondary antibody
Indirect immunofluorescence
39
Which technique uses antigen-antibody interaction to highlight the presence of antigens rather than tissue structures
Immunohistochemistry + Immunofluorescence
40
Which technique is used for targeted therapy in cancer
Immunohistochemistry
41
Which technique is used for diagnosis of autoimmune disorders
Immunofluorescence
42
Which reaction is used in immunohistochemistry with chromogen and enzyme
Immuno-enzymatic reaction
43
Which feature makes chromogenic products easy to detect but difficult to quantitate
Colored precipitate is easy to detect but hard to quantitate
44
Which technique is more sensitive than immunofluorescence
Immunohistochemistry
45
Which microscope is used for immunohistochemistry
Light microscope
46
Which step in immunohistochemistry requires more time
Incubation and blocking steps
47
Which property allows chromogenic slides to be stored for years
Colored precipitate is not sensitive to light
48
Which color is most common in immunohistochemistry
Brown
49
Which chromogen is most commonly used in immunohistochemistry
DAB
50
Which enzyme is most commonly used in immunohistochemistry
Horseradish peroxidase
51
Which counterstain is most commonly used in immunohistochemistry
Hematoxylin
52
Which step is first in chromogenic method
Sample preparation
53
Which step follows sample preparation in chromogenic method
Labeling
54
Which tissue type was originally used for immunofluorescence
Fresh tissue
55
Which tissue type is now commonly used for immunohistochemistry
Paraffin fixed embedded tissue
56
Which step is needed for paraffin embedded tissue before immunohistochemistry
Pretreatment or antigen retrieval
57
Why is antigen retrieval necessary
Formalin crosslinking masks antigens
58
Which step follows antigen retrieval
Protein blocking
59
Why is protein blocking performed
To prevent false positive reactions from non-target proteins
60
Which reagent is added after protein blocking
Primary antibody
61
Which reagent is added after primary antibody
Secondary antibody
62
Which reagent is added after secondary antibody
Streptavidin peroxidase
63
Which reagent is added after streptavidin peroxidase
DAB chromogen
64
Which step follows DAB chromogen
Counterstaining with hematoxylin
65
Which factors affect the degree of antigen masking
Length of time in fixative + Temperature + Concentration of fixative + Availability of other proteins for cross-linkage
66
What is the ideal fixation time for immunohistochemistry
22–72 hours
67
Which enzymes are used for proteolytic digestion
Trypsin + Protease
68
What is the effect of proteolytic digestion
Breaks down formalin cross-linking and exposes antigenic sites
69
Which antigens are commonly demonstrated by proteolytic digestion
Heavy chain immunoglobulins + Complement + Specific antigens like CK
70
What happens with under-digestion
Too little staining
71
What happens with over-digestion
False positive staining + High background staining + Tissue damage
72
Which retrieval method uses heat and buffered solutions
Heat-induced epitope retrieval
73
What does heat-induced epitope retrieval do
Breaks crosslinks and removes bound calcium ions
74
Which solutions are used for microwave heat-induced epitope retrieval
0.01 M sodium citrate buffer at pH 6.0 + 0.1 mM EDTA at pH 8.0
75
Which device is used for pressure cooker heat-induced epitope retrieval
Pressure cooker at 15 psi and 120 degrees Celsius
76
Which devices are used for steamer and water bath heat-induced epitope retrieval
Steamer + Water bath at 95–98 degrees Celsius
77
Which advantage is associated with heat-induced epitope retrieval
Many antigens are retrieved regardless of fixation length
78
Which advantage is associated with heat-induced epitope retrieval for heavy chain immunoglobulins
More reliable and reproducible demonstration
79
Which disadvantage is associated with heat-induced epitope retrieval
Drying destroys antigenicity
80
Which disadvantage is associated with heat-induced epitope retrieval
Boiling damages nuclear detail
81
What is non-specific staining in immunohistochemistry
Binding of primary antibody to amino acids other than the desired epitope
82
Why is high background staining a problem
Masks detection of target antigen
83
Which sources contribute to non-specific staining
Endogenous enzymes or fluorochromes + Endogenous biotin + Endogenous antibody binding activity + Cross reactivity of secondary reagents with endogenous proteins
84
Where is endogenous peroxidase found
Leukocytes + Erythrocytes
85
How is endogenous peroxidase blocked
Incubation in absolute methanol containing 0.5 percent hydrogen peroxide for 10 minutes at room temperature
86
Where is endogenous alkaline phosphatase found
Lymphoid tissue + Intestinal tissue + Placenta
87
How is endogenous alkaline phosphatase blocked
1 nM Levamisole
88
Where are hydrophobic interactions causing non-specific staining found
Collagen + Connective tissues + Epithelium + Adipocytes
89
How are hydrophobic interactions blocked
Addition of blocking protein Triton X or high salt concentration 2.5 percent NaCl
90
What causes primary antibody to bind non-immunologically to highly charged groups
Highly charged groups in connective tissues
91
How is non-immunological binding to highly charged groups blocked
Addition of protein solution casein to saturate and neutralize charged sites
92
Which molecules are attached to antibodies for visualization
Labels
93
Which types of antibodies are used as primary reagents
Polyclonal antibodies + Monoclonal antibodies
94
Which type of antibody is produced by multiple clones
Polyclonal antibodies
95
Which type of antibody is produced by a single clone
Monoclonal antibodies
96
Which animal is commonly used for polyclonal antibody production
Rabbit
97
Which cells are activated in polyclonal antibody production
B cells
98
Which cells produce antibodies in polyclonal antibody production
Plasma cells
99
Which technology is used to produce monoclonal antibodies
Hybridoma technology
100
Which animal is commonly used for monoclonal antibody production
Mouse
101
Which cells are fused with antibody-producing plasma cells in hybridoma technology
Cancerous plasma cells (myeloma cells)
102
Why are myeloma cells used in hybridoma technology
Because they are immortal and allow continuous cell culture
103
What is the result of hybridoma technology
Individual hybridoma plasma cells
104
Which antibody type is more specific for immunohistochemistry
Monoclonal antibodies
105
Which antibody type can react with multiple antigens
Polyclonal antibodies
106
Which labels are used for immunohistochemistry
Enzyme labels + Fluorescent labels + Radiolabels
107
Which label is most widely used in immunohistochemistry
Enzyme labels
108
Which label uses chromogens for detection
Enzyme labels
109
Which label uses fluorochromes for detection
Fluorescent labels
110
Which label uses radioisotopes for detection
Radiolabels
111
Which enzyme is most commonly used in immunohistochemistry
Horseradish peroxidase
112
Which chromogen is most commonly used in immunohistochemistry
3
113
What is the product of horseradish peroxidase and DAB
Insoluble stable dark brown reaction product
114
Which microscope is used for enzyme-labeled immunohistochemistry
Light microscope
115
Why is the product of HRP and DAB suitable for long storage
It is not light sensitive
116
Which counterstain is most commonly used in immunohistochemistry
Hematoxylin
117
What is the purpose of counterstaining
To stain the nucleus
118
What are other chromogens used in immunohistochemistry
3-amino-9-ethylcarbazole + 4-chloro-1-naphthol + Hanker-Yates reagent + A-naphthol pyronin + Vector Red + Vector Blue + Vector VIP + BCIP/NBT + Calf intestinal alkaline phosphatase + Fast red TR with naphthol AS-MX phosphate sodium salt
119
Which method uses calf intestinal alkaline phosphatase
APAAP method
120
Which chromogen produces a bright red reaction product
Fast red TR with naphthol AS-MX phosphate sodium salt
121
Which labels are used for immunofluorescence
Fluorochromes
122
What happens when fluorochromes absorb light
Electrons reach an unstable excited state
123
What happens when electrons return to ground state in fluorochromes
Emit light of a different longer wavelength
124
Is fluorescence in immunofluorescence permanent
No
125
Which tissue type is most commonly used for immunofluorescence
Fresh tissue
126
Which tissue type is most commonly used for immunohistochemistry
Formalin-fixed paraffin-embedded tissue
127
Which fluorochromes are most widely used
Fluorescein isothiocyanate + Tetramethyl rhodamine isothiocyanate
128
What is the emission color of FITC
Apple-green
129
What is the emission color of TRITC
Orange-red
130
Which absorption spectrum do FITC and TRITC have
Ultraviolet to blue light
131
How are FITC and rhodamine linked to antibodies
Covalently to free amino groups on lysine side chains and free carboxyl groups in aspartic acid and glutamic acid residues
132
At what pH does the reaction occur for FITC and rhodamine conjugation
pH 9.5
133
Is the conjugation of fluorochromes to antibodies time and temperature dependent
Yes
134
What is the optimal fluorochrome to antibody ratio
2 to 4:1
135
What is the effect of over-conjugation
High background staining
136
What is the effect of under-conjugation
Unsatisfactory low-level fluorescence
137
How is free chromophore removed from conjugate preparation
Dialysis against 0.15 M sodium chloride at 4 degrees Celsius + Gel infiltration column chromatography using Sephadex G50
138
Which blocking techniques are used in immunohistochemistry
Levamisole + Casein
139
Can immunofluorescence also have non-specific staining
Yes
140
What is free chromophore
Fluorochrome not conjugated to antibody
141
Which labels require autoradiographic facilities for detection
Radioisotopes
142
Which labels are not used in routine diagnostic laboratories
Radioisotopes
143
Which quality control component is used in immunohistochemistry and immunofluorescence
Controls
144
Which control is known and proven to contain the antigen in question
Positive control
145
Which control uses a tissue section known to express the antigen
Positive control
146
Which control is placed on the same slide as the patient sample
Positive control
147
Which staining pattern is expected in estrogen and progesterone receptor assays
Nuclear staining
148
Which staining pattern is expected in HER2NEU assays
Complete membranous staining
149
Which control uses a parallel section with an unrelated immunoglobulin instead of the primary antibody
Negative control
150
Which control ensures true negative results by providing a positive reference
Positive control
151
Which control is built into the tissue itself and does not require an additional section
Internal tissue control
152
Which control contains the target antigen in adjacent normal tissue
Internal tissue control
153
Which example demonstrates a strong internal control in breast cancer
Normal ductal epithelium positive and tumor negative
154
Which example indicates a need to repeat the procedure due to negative internal control
Both normal and tumor negative
155
Which example suggests poor staining quality
Both normal and tumor weakly positive
156
Which method labels the primary antibody directly with an enzyme or fluorochrome
Direct technique
157
Which method demonstrates immunoglobulin and complement in frozen sections
Direct technique
158
Which method is quick and easy but provides little signal amplification
Direct technique
159
Which method uses a primary antibody labeled with horseradish peroxidase
Direct technique
160
Which method uses a primary antibody labeled with a fluorochrome
Direct technique
161
Which technique is used for immunohistochemistry with enzyme labels
Direct technique
162
Which technique is used for immunofluorescence with fluorochrome labels
Direct technique
163
Which method attaches primary antibody and peroxidase enzyme to a dextran polymer backbone
Enhance polymer one-step staining method
164
Which method reduces the number of incubation steps
Enhance polymer one-step staining method
165
Which method is more sensitive
Enhance polymer one-step staining method
166
Which method is suitable for frozen immunohistochemistry
Enhance polymer one-step staining method
167
Which method eliminates non-specific staining from endogenous biotin
Enhance polymer one-step staining method
168
Which method has a limited number of commercially available primary antibodies
Enhance polymer one-step staining method
169
Which method uses a labeled secondary antibody directed against an unlabeled primary antibody
Indirect technique
170
Which method is more sensitive than direct technique
Indirect technique
171
Which method allows multiple secondary antibodies to react with different sites on the primary antibody
Indirect technique
172
Which method uses an unconjugated primary antibody and a secondary antibody conjugated to an enzyme-labeled polymer chain
Polymer chain two-step indirect technique
173
Which method is quick reliable and sensitive
Polymer chain two-step indirect technique
174
Which method is most commonly used in routine diagnostic immunohistochemistry
Polymer chain two-step indirect technique
175
Which method uses a soluble peroxidase-antiperoxidase complex bound to an unconjugated primary antibody by a bridging antibody
Peroxidase-antiperoxidase technique
176
Which method produces an insoluble dark brown reaction product
Peroxidase-antiperoxidase technique
177
Which blocking technique is used in peroxidase-antiperoxidase technique
Absolute methanol containing hydrogen peroxide
178
Why is blocking used in peroxidase-antiperoxidase technique
To block endogenous peroxidase and prevent background staining
179
Which technique avoids interference from endogenous peroxidase activity
Alkaline phosphatase-anti-alkaline phosphatase complex
180
Which specimens are ideal for alkaline phosphatase-anti-alkaline phosphatase technique
Blood + Bone marrow smears
181
How is endogenous alkaline phosphatase activity blocked
By adding levamisole to the substrate solution
182
Which enzyme complex binds with the primary antibody using a secondary antibody as a bridge
Alkaline phosphatase-anti-alkaline phosphatase complex
183
Why is blocking needed in alkaline phosphatase-anti-alkaline phosphatase technique
To prevent background staining from endogenous alkaline phosphatase
184
Which technique uses an unconjugated primary antibody
biotinylated secondary antibody
185
Which enzyme is used in the avidin-biotin technique
Horseradish peroxidase + Alkaline phosphatase
186
Which chromogen is used in the avidin-biotin technique
Chromogen
187
Which technique is called LSAB
Labelled streptavidin-biotin technique
188
Which technique is called ABC
Avidin-biotin complex technique
189
How long is the avidin-biotin complex prepared before use
30 minutes
190
At what temperature is the avidin-biotin complex incubated
Room temperature
191
What is the sequence of steps in the avidin-biotin technique
Primary antibody + Biotinylated secondary antibody + Avidin-biotin complex
192
Which property of avidin allows strong binding to biotin
High affinity for biotin
193
Which disadvantage is associated with avidin
High isoelectric point and positive charge at neutral pH
194
Which disadvantage is associated with avidin
Tendency to react with lectins via carbohydrate moiety
195
Which tissues require avidin/biotin block before primary antibody
Liver + Kidney
196
Which technique is 4–8 times more sensitive than ABC
LSAB
197
Which step is first in immunohistochemistry
Tissue sections from paraffin embedded tissue block
198
Which step follows tissue sectioning
Antigen retrieval
199
Which step follows antigen retrieval
Blocking endogenous enzymes
200
Which step follows blocking endogenous enzymes
Blocking background staining
201
Which step follows blocking background staining
Primary antibody
202
Which step follows primary antibody
Secondary antibody
203
Which step follows secondary antibody
Chromogen substrate
204
Which step follows chromogen substrate
Counterstain
205
Which step follows counterstain
Mounting
206
Which step follows mounting
Microscopic examination
207
Which microscope is used for final examination
Light microscopy
208
Which marker is used for epithelial cells
Keratin
209
Which specific keratin is positive in lung
breast
210
Which specific keratin is positive in colon and stomach carcinoma
CK20
211
Which tumors are CK7+/CK20+
Urothelial carcinoma + Mucinous ovarian tumors
212
Which tumors are CK7-/CK20-
Renal cell carcinoma + Thyroid carcinoma + Squamous cell carcinoma
213
Which marker is a transmembrane protein in glandular or luminal epithelial cells
Epithelial membrane antigen
214
Which tumors are positive for epithelial membrane antigen
Adenocarcinoma of breast + Lung + Kidney
215
Which tumors are non-reactive for epithelial membrane antigen
Hepatocellular carcinoma + Adrenal carcinoma + Embryonal carcinoma
216
Which tumors are negative for epithelial membrane antigen
Sarcoma + Lymphoma + Melanoma
217
Which oncofetal antigen is present in carcinomas of the gastrointestinal tract
pancreas
218
Which tumor is positive for carcinoembryonic antigen
Adenocarcinoma
219
Which tumor is negative for carcinoembryonic antigen
Mesothelioma
220
Which marker distinguishes lung adenocarcinoma from mesothelioma
Thyroid transcription factor-1
221
Which tumors are positive for thyroid transcription factor-1
Thyroid + Lung + Neuroendocrine tumors
222
Which marker is positive in prostatic adenocarcinoma
Prostate specific antigen
223
Which other tumors can be positive for prostate specific antigen
Pancreatic tumor + Salivary gland tumor
224
Which biopsy result confirms metastatic prostatic adenocarcinoma
Positive PSA in iliac lymph node
225
What is the difference between non-reactive and negative in epithelial membrane antigen
Non-reactive is carcinoma but does not stain
226
Which stains are used first in tumor diagnosis
H&E
227
Which step is done before immunohistochemistry in tumor diagnosis
Morphologic assessment with H&E
228
Which contractile intermediate filament protein is present in muscle and some non-muscle tissue
Actin
229
Which marker is sensitive for muscle differentiation
Actin
230
Which tumors are identified by actin as a marker
Tumors derived from smooth + Skeletal + Cardiac muscle
231
Which tumor shows diffuse and strong cytoplasmic positivity for SMA
Leiomyoma
232
Which tumor is similar to leiomyoma in SMA positivity
Leiomyosarcoma
233
Which intermediate filament is expressed by smooth and striated muscle
Desmin
234
Which tumors are positive for desmin
Leiomyoma + Rhabdomyosarcoma
235
Which cells show strong diffuse cytoplasmic positivity for desmin in rhabdomyosarcoma
Rhabdoid cells
236
Which intermediate filament protein is expressed by central nervous system glial cells
Glial fibrillary acidic protein
237
Which cell type specifically expresses glial fibrillary acidic protein
Astrocytes
238
Which tumors are positive for glial fibrillary acidic protein
Astrocytoma + Ependymoma + Oligodendroglioma + Medulloblastoma
239
Which tumors are negative for glial fibrillary acidic protein
Meningioma + Metastatic carcinoma + Lymphoma
240
Where is glial fibrillary acidic protein strongly positive in ependymoma
Perivascular processes
241
Which intermediate filament is expressed in neural origin cells
Neurofilament
242
Which cell types express neurofilament
Neurons + Neuronal processes + Peripheral nerves + Sympathetic ganglia + Adrenal medulla + Neuroendocrine cells
243
Which tumors are positive for neurofilament
Neuroblastoma + Ganglioneuroma + Neuroma + Pheochromocytoma
244
Which calcium-binding protein is expressed in central nervous system glial cells
Schwann cells
245
Which diagnosis is confirmed by S-100 positivity
Melanoma
246
Which marker is an isoenzyme for neural or neuroendocrine differentiation
Neuron-specific enolase
247
Which tumor is identified by neuron-specific enolase
Pheochromocytoma
248
Which marker is found in neural secretory granules of endocrine tissues
Chromogranin
249
Which staining pattern is typical for chromogranin
Granular
250
Which tumor is positive for both chromogranin and cytokeratin
Neuroendocrine carcinoma
251
Which tumor is positive for chromogranin but negative for keratin
Paraganglioma
252
Which transmembrane protein is associated with presynaptic vesicles of neurons
Synaptophysin
253
Which cells express synaptophysin
Normal neurons + Neuroendocrine cells
254
Which markers are used for neuroendocrine tumors
CD56 + Synaptophysin + Chromogranin
255
Which tumor has a low proliferation index and is positive for neuroendocrine markers
Pulmonary typical carcinoid
256
Which marker is produced by placental syncytiotrophoblasts and used for choriocarcinoma
Human chorionic gonadotropin
257
Which cells produce beta-hCG
Syncytiotrophoblasts
258
Which tumor is associated with malignant trophoblastic proliferation
Choriocarcinoma
259
Which marker is synthesized by normal liver hepatocytes
Alpha-fetoprotein
260
Which tumor marker is used for endodermal sinus tumors with yolk sac differentiation
Alpha-fetoprotein
261
Which tumors can also be positive for alpha-fetoprotein
Embryonal carcinoma + Teratoma + Hepatocellular carcinoma
262
Which marker is produced by placental syncytiotrophoblasts in late pregnancy
Placenta-like alkaline phosphatase
263
Which tumors are marked by placenta-like alkaline phosphatase
Germinoma + Embryonal carcinoma + Choriocarcinoma + Endodermal sinus tumor + Seminoma
264
Which tumors are considered muscle specific tumors
Myogenic tumors
265
Which markers are used for muscle specific tumors
Muscle specific actin + Desmin + Myo-D1 + Myoglobin + Myogenin
266
Which tumor is a malignant tumor of smooth muscle origin
Leiomyosarcoma
267
Which marker is positive in leiomyosarcoma using immunohistochemistry
Desmin
268
Which markers are used for fibrohistiocytic tumors
CD68 + FAM 56
269
Which tumor is an example of fibrohistiocytic tumor
Reticulohistiocytoma of the skin
270
Which markers are used for vascular tumors
Factor VIII-related antigen + CD31 + Ulex Europaeus 1
271
Which tumor shows strong vascular marker expression on CD31 immunostain
Cutaneous angiosarcoma
272
Which tumor is derived from melanocytes
Melanoma
273
Which cells are derived from the neural crest and react with S100
Melanocytes
274
Which marker is highly sensitive and specific for melanoma
Melanosome HMB-45
275
Which marker is present in normal and pigmented cells of skin
retina
276
Which markers are used for melanoma
S100 + Melanosome HMB-45 + Melan-A MART-1
277
Which diagnostic method is useful for lymphoma diagnosis
Immunohistochemistry
278
Which marker is the best screening marker for lymphoma
LCA CD45
279
Which marker distinguishes lymphoma from carcinoma
CD45
280
Which markers are used for T-cell lymphomas
CD3 + CD4 + CD5
281
Which markers are used for B-cell lymphomas
CD19 + CD20 + CD23
282
Which markers are used for Hodgkin lymphoma Reed-Sternberg cells
CD15 + CD30
283
Which staining pattern is seen for CD30 in Reed-Sternberg cells
Membrane and cytoplasm diffusely and weakly
284
Which staining pattern is seen for CD15 in Reed-Sternberg cells
Crisp membrane staining and Golgi staining
285
Which markers can detect tumors with high or low proliferative activity
Cell proliferation markers
286
Which marker is most commonly used for cell proliferation
Ki-67 MIB-1
287
Which marker is also used for cell proliferation
Proliferating cell nuclear antigen
288
Which cells are negative for proliferation markers
Non-proliferating cells
289
Which cells are positive for proliferation markers
Proliferating cells
290
How is Ki-67 reported in tumor evaluation
Percentage of tumor cells positive
291
Which factors are considered in grading tumors with Ki-67
Percentage and intensity of staining
292
Which cancer-associated genes are commonly analyzed
p53 + C-erbB-2 + C-myc
293
Which infectious agent marker is detected in liver sections by immunohistochemistry
Hepatitis B virus surface antigen
294
Which infectious agent marker is associated with cervical cancer
HPV
295
Which infectious agent marker can be associated with cervical cancer independently of HPV genotype
p16
296
Which infectious agent markers are used for infectious agent detection
Hepatitis virus + HPV + EBV + Mycobacteria + Cryptococcus neoformans + H. pylori
297
Which surrogate marker is used for HPV driven squamous cell carcinoma
p16
298
Which staining pattern is seen for p16 in high grade squamous epithelial lesions
Diffuse and strong nuclear and cytoplasmic staining
299
How is the grading of immunohistochemistry results for p16 performed
By percentage and intensity of staining
300
Which technique generates high-resolution images for protein localization
Immunofluorescence
301
Which technique is used to quantitate fluorescent signal
Immunofluorescence
302
Which diseases are diagnosed using immunofluorescence
Glomerular diseases + Systemic lupus + Vasculitis
303
Which samples can be evaluated by immunofluorescence
Cells in suspension + Cultured cells + Tissue + Beads + Microarrays
304
Which types of samples can be analyzed for specific proteins using immunofluorescence
Fresh + Fixed samples
305
Which methods are used for immunofluorescence
Direct method + Indirect method
306
Which direct immunofluorescence method uses antibodies chemically conjugated to fluorescent dyes
Direct labeling
307
Which samples are used for direct immunofluorescence
Cryostat section of fresh 2–5 mm thin tissue + Fresh and fixed material mounted on slides coated with gelatin adhesive or poly-L-lysine at 1:10 dilution
308
Which antibody binds directly to the antigen in direct immunofluorescence
Primary antibody tagged with fluorophore
309
Which method uses a primary antibody followed by a secondary antibody conjugated to a fluorophore
Indirect immunofluorescence
310
Which technique is used for detection of autoantibodies in patient serum
Indirect immunofluorescence
311
Which autoantibodies are detected by indirect immunofluorescence
Anti-nuclear antibodies + Anti-mitochondrial antibodies + Microsomal antibodies
312
Which technique is used for direct labeling of leukemia cells
Direct immunofluorescence
313
Which color is commonly used for direct labeling of leukemia cells
Green
314
Which pattern is seen in anti-nuclear antibody immunofluorescence
Diffuse patterns depending on the antigens detected
315
What is observed in H&E for capillary walls in immune complex diseases
Thick capillary walls
316
What is observed in immunofluorescence microscopy for capillary walls in immune complex diseases
Immune complex deposits in capillary walls
