Study Guide: Neuroanatomical Techniques Flashcards
1
Q
Nissl Stain developed by:
A
Franz Nissl
2
Q
Nissl Stain is used to:
A
differentiate neurons and glial cells, study neuronal shapes and sizes
3
Q
Nissl stain uses what kind of dye?
A
Basic aniline dyes (e.g. cresyl violot, toluidine blue, methylene blue)
4
Q
The basic aniline dye used by Nissl stain binds to
A
negatively charged nucleic acids (RNA and DNA)
5
Q
Nissl stain stains nuclei and Nissl bodies, the latter which is:
A
Rough ER with with ribosomes and nuclei
6
Q
Nissl stain highlights:
A
neurons over glial cells due to higher RNA concentration (neurons have a much higher protein synthesis activity)
7
Q
Nissl Stain: Application
A
Used for:
NEURONAL CYTOARCHITECTURE studies and examining
NEURONAL POPULATION CHANGES
8
Q
Golgi stain was developed by:
A
Camille Golgi in the 1870s
9
Q
Golgi stain mechanism:
A
- Fixation
- Impregnation
10
Q
Describe the fixation mechanism (first step) of Golgi stains:
A
Tissue is treated with potassium dichromate to create reactive sites
11
Q
Describe the impregnation mechanism (step #2) of Golgi stains:
A
Impregnation: silver nitrate (AgNO3) reacts with dichromate-treated tissue forming silver chromate microcrystals
12
Q
Golgi stain: selective staining:
A
Only a subset of neurons are stained, allowing detailed visualisation of dendrites and axons
13
Q
Golgi stain: Applications(2):
A
(1) Used extensively by Santiago Ramon y Cajal to develop the neuron doctrine
(2) Visualizing dendritic spines and neuronal morphology
14
Q
Nissle stain: Target
A
Nissle bodies (rough Er)
15
Q
Golgi stain: Target:
A
Entire neuron (soma, dendrites, axons)
16
Q
Nissl Stain: Selectivity
A
Labels all neurons
17
Q
Golgi Stain: Selectivity:
A
Labels only a subset of neurons
18
Q
Nissl Stain Primary use:
A
Cytoarchitecture
19
Q
Golgi Stain: Primary use:
A
Morphology of individual neurons
20
Q
Nissle Stain Color:
A
Blue/purple
21
Q
Golgi stain color:
A
Black
22
Q
Immunocytochemistry (ICC) / Immunohistochemistry (IHC)
A
Uses antibodies to detect specific proteins (e.g. neurotransmitters, receptors)
23
Q
Immunocytochemistry (ICC) / Immunohistochemistry (IHC): Applications (3) :
A
(1) Identifying neurons vs glia
(2) Mapping neurotransmitter distribution
(3) Studying pathological conditions (e.g. neurodegeneration, tumors)
24
Q
Immunocytochemistry (ICC) / Immunohistochemistry (IHC): mechanism:
A
(1) Antibodies bind to target proteins (antigens)
(2) Markers used for visualizations
25
What are the two common types of markers used for visualization inImmunocytochemistry (ICC) / Immunohistochemistry (IHC): What are the two types of markers used for visualization:
(1) Fluorescent dyes --> fluorescence microscopy
(2) Enzymes (horseradish peroxidase) --> colored reaction under light microscopy
26
Advantages of immunocytochemistry/ immunohistochemistry (2) :
-High specificity and sensitivity
-Can be combined with confocal microscopy for 3D imaging
27
Limitations (2) of immunocytochemistry / immunohistochemistry:
(1) Requires well -validated antibodies
(2) Signal strength varies with antigen accessibility
28
In situ hybridization (ISH):
Uses complementary RNA probes to detect mRNA expression
29
Applications In situ hybridization (ISH) (2):
(1) Research: Gene expression mapping in neurons
(2) Clinical: Brain tumour diagnosis, schizophrenia research (identifying altered transcript localization)
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In situ hybridization (ISH): procedure:
1. Fixation
2. Probe design
3. Hybridization
4. Washing
5. Detection
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In situ hybridization: Fixation
Preserves tissue structure
32
In situ hybridization: Probe design:
Complementary to target mRNA
32
In situ hybridization: Hybridization:
Probe binds to mRNA
33
In situ hybridizaiton: Washing:
Removes unbound probes
34
In situ hybridization: Detection:
Fluorescence, autoradiography, or enzyme reactions
35
In situ hybridizaiton: Advantages:
High-resolution gene expression mapping
36
In situ hybridization: limitations:
Time - intensive, requires careful probe design
37
What are two genetic techniques (2):
(1) Brainbow techniques
(2) Optogenics
38
Brainbow technique uses:
genetically encoded fluorescent proteins to label neurons in multiple colors
39
Brainbox technique: Applications (2):
(1) Single cell resolution mapping of neural circuits. Each cell is labelled with a unique color, so that neurites can be individually traced (e.g. you can find out where a specific axon goes in order to understand how the circuit is connected)
(2) DNA constructs with fluorescent proteins (GFP,CFP,YFP) are randomly recombined, producing unique neuron colors
40
Optogenics uses:
Light sensitive ion channels (e.g. channelrhodospins) to control neuronal activity with light
41
Optogenics applications(2):
(1) Mapping neuronal connectivity (e.g. whether a set of neurons in area A excites or inhibits a downstream neuronal poopulation in area B)
(2) Understanding behaviour- related neuronal activity
42
Viral based tracing:
Uses modified viruses to trace synaptic connections
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Viral based tracing: common tracers:
(1) herpes simplex virus (Multi synaptic tracing)
(2) Rabies virus (retrograde transsynaptic tracing)
(3) Pseudorabies virus (PRV) (traces from axon terminals to soma)
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Viral based tracing: Herpes simplex virus:
Multi synaptic tracing
45
Viral based tracing: rabies virus:
retrograde transsynaptic tracing
46
Viral based tracing: Pseudorabies virus (PRV) :
traces from axon terminals to soma
47
Viral based tracing: Applications (2):
(1) Mapping functional networks
(2) Studying long-range neural pathways
48
Dye based tracing:
uses axonal transport to trace connections
49
Dyes used in Dye based tracing:
Horseradish peroxidase (HRP)
Fluorogold
Dextrans
50
Transport types in Dye -Based tracing:
Anterograde: Soma -> axon terminals
Retrograde: Axon terminals -> Soma
51
Diffusion tensor imaging:
specialized MRI that maps white matter tracts
52
Diffusion Tensor Imaging (DTI): Mechanism:
Specialized MRI that maps white matter tracts
53
Diffusion tensor imaging: mechanism:
Tracks anisotropic water diffusion along axons
54
Diffusion tensor imaging (DTI): Applications (2):
(1) Mapping brain connectivity
(2) Detecting axonal damage
55
Computational neuroanatomy:
combines imaging techniques and computational tools for modeling and analysis
56
Computational neuroanatomy:
Combines imaging techniques and computational tools for modeling and analysis
57
Computational neuroanatomy: applications (3):
(1) disease diagnosis
(2) Studying brain plasticity and learning
(3) machine learning for neural structure
58
1. Which of the following is not a neuroanatomical technique?
a) Histological techniques
b) Electrophysiology
c) Non-invasive imaging
d) Dye-based tracing
B) Electrophysiology
59
2. Who developed the Nissl stain?
a) Santiago Ramón y Cajal
b) Camillo Golgi
c) Franz Nissl
d) Heinrich Waldeyer
Frank Nissl
60
3. What is the primary target of Nissl staining?
a) Axon terminals
b) Myelin sheath
c) Rough endoplasmic reticulum (Nissl bodies)
d) Astrocyte processes
c) Rough endoplasmic reticulum (Nissl bodies)
61
4. What type of dye is used in Nissl staining?
a) Acidic aniline dyes
b) Basic aniline dyes
c) Fluorescent dyes
d) Silver nitrate
b) Basic aniline dyes
62
5. What is a Nissl body?
a) Cluster of mitochondria
b) Synaptic vesicle storage site
c) Aggregation of rough endoplasmic reticulum
d) Part of the Golgi apparatus
c) Aggregation of rough endoplasmic reticulum
63
6. The Golgi stain was developed by:
a) Franz Nissl
b) Camillo Golgi
c) Santiago Ramón y Cajal
d) Rudolf Virchow
b) Camillo Golgi
64
7. What chemical is first used in the Golgi staining method?
a) Silver nitrate
b) Potassium dichromate
c) Fluorogold
d) Cresyl violet
b) Potassium dichromate
65
8. Which reaction forms the black deposits in Golgi staining?
a) Silver nitrate reacting with cell membrane proteins
b) Potassium dichromate reacting with lipids
c) Silver chromate precipitation
d) Fluorescence of bound dyes
c) Silver chromate precipitation
66
9. What is the major advantage of Golgi staining?
a) Selectively stains entire neurons
b) Shows differences between glia and neurons
c) Labels only cell nuclei
d) Identifies myelin proteins
a) Selectively stains entire neurons
67
10. Santiago Ramón y Cajal used the Golgi stain to:
a) Discover the synapse
b) Provide evidence for the neuron doctrine
c) Identify neurotransmitters
d) Classify glial cells
b) Provide evidence for the neuron doctrine
68
11. What is the purpose of immunocytochemistry (ICC)?
a) To detect and visualize proteins using antibodies
b) To identify glial cell morphology
c) To analyze myelin thickness
d) To perform non-invasive imaging
a) To detect and visualize proteins using antibodies
69
12. Which labeling methods are commonly used in immunocytochemistry ICC?
a) Fluorescent dyes
b) Silver nitrate staining
c) Aniline dyes
d) Golgi silver chromate
a) Fluorescent dyes
70
13. In situ hybridization (ISH) is used to:
a) Detect protein expression in neurons
b) Detect specific mRNA sequences (which reveals that the neuron synthesizes the protein/peptide encoded by that mRNA sequence)
c) Trace neuronal connections
d) Identify axonal transport pathways
b) Detect specific mRNA sequences (which reveals that the neuron synthesizes the protein/peptide encoded by that mRNA sequence)
71
14. Brainbow technology is used to:
a) Trace neural circuits using viral markers
b) Genetically label neurons with different fluorescent proteins
c) Visualize synaptic vesicles
d) Measure neurotransmitter levels
Genetically label neurons with different fluorescent proteins
72
15. The Brainbow technique relies on:
a) Random recombination of fluorescent proteins
b) Silver chromate staining
c) Immunohistochemistry
d) Axonal transport
a) Random recombination of fluorescent proteins
73
16. Optogenetics allows researchers to:
a) Control neurons with light
b) Track neurotransmitter release
c) Measure synaptic potentials
d) Detect protein interactions
a) Control neurons with light
74
17. Viral tracing techniques help:
a) Map synaptic connectivity
b) Detect neurotransmitter levels
c) Identify gene expression
d) Analyze action potentials
Map synaptic connectivity
75
18. Which virus is commonly used for retrograde transsynaptic tracing?
a) Herpes simplex virus (HSV)
b) Rabies virus
c) Influenza virus
d) Poliovirus
b) Rabies virus
76
19. Which dye-based method is commonly used for neuronal tracing?
a) Horseradish peroxidase (HRP)
b) Brainbow
c) Golgi stain
d) MRI
Horseradish peroxidase (HRP)
77
20. Anterograde transport refers to:
a) Movement from axon terminals to soma
b) Movement from soma to axon terminals
c) Diffusion of dyes across synapses
d) Viral transmission between neurons
) Movement from soma to axon terminals
78
21. What does MRI primarily measure?
a) Neuronal firing rates
b) Magnetic properties of hydrogen nuclei
c) Glucose metabolism
d) Electrical activity
b) Magnetic properties of hydrogen nuclei
79
22. What type of MRI technique maps white matter tracts?
a) Functional MRI (fMRI)
b) Diffusion Tensor Imaging (DTI)
c) Magnetoencephalography (MEG)
d) Golgi staining
b) Diffusion Tensor Imaging (DTI)
80
23. DTI imaging relies on:
a) Diffusion of water molecules
b) Radioactive tracers
c) Ion channel activity
d) Myelin protein expression
a) Diffusion of water molecules
81
24. A voxel in MRI refers to:
a) A measure of brain volume
b) The smallest unit of 3D spatial resolution
c) A type of contrast agent
d) A synaptic marker
b) The smallest unit of 3D spatial resolution
82
26. Which of the following are limitations of MRI?
a) Can’t image white matter
b) Expensive and contraindicated for metal implants and pace-makers
c) Uses radioactive tracers
d) Requires invasive surgery
Expensive and contraindicated for metal implants and pace-makers
83
27. Computational neuroanatomy combines:
a) Imaging and computational modelling tools
b) Golgi and Nissl staining
c) Immunocytochemistry and Brainbow
d) Dye-based and viral tracing
Imaging and computational modelling tools
84
28. What is a major application of computational neuroanatomy?
a) Diagnosing neurodegenerative diseases
b) Staining neurons
c) Measuring neurotransmitter levels
d) Axonal transport studies
Diagnosing neurodegenerative diseases
85
