RESEARCH METHODS IN
NEUROSCIENCE EXAM QUESTIONS
AND ANSWERS
Ways to Fix Tissue - Answer-Immersion- place entire specimen in fixative solution
Perfusion- deliver fixative through cardiovascular system
Process of Histology: - Answer-Fixation, embedding, sectioning
Embedding - Answer-Specimens sometimes embedded in gelatin, plastic, par
paraffin wax; stabilizes structure to make it easier to cut
Sectioning - Answer-Cutting of tissue into thin slices using sliding microtome, rotary
microtome, cryostat, vibratome
Sliding microtome - Answer-Frozen, blade moves
Rotary microtome - Answer-Not frozen, in plastic
Cryostat - Answer-Blade is fixed, specimen moves (inverse of microtome), frozen
Vibratome - Answer-Slice fresh tissue, vibrating knife
Why non-fixed tissue? - Answer-If fixative may damage or prevent histological
procedure, can freeze to prevent decay
3 Orientations to cut brain - Answer-Coronal, sagittal, horizontal
Types of Stains: - Answer-Cell body, fiber, golgi
Cresyl Violet - Answer-Cell body stain, stains cell nuclei and nissel, blue to purple,
useful for examining of cytoarchitecture, stains each type of neuron slightly
differently
Hematoxylin - Answer-Cell body stain, stains cell nuclei, blue to blue-black, often
used in combination with Eosin
Eosin Y - Answer-Stains cytoplasm, pink to red
Thionine - Answer-Cell body stain, stains cell nuclei and nissel stain, blue to purple
Weigert - Answer-Fiber stain, stains myelin, normal myeline is blue, degenerated
myelin is yellow, combines with hematoxylin to selectively stain myelin
Golgi stain - Answer-Fills neuron cell bodies and processes, black, stains individual
neurons at random
, Cell body stains- basophilic stains - Answer-Stains acidic molecules like RNA and
DNA (see dendrites, nucleolus, nissel substance, axon hillock, NOT axon
In Situ Hybridization - Answer-Shows where and when a gene is transcribed (mRNA)
1. ID mRNA sequence to be studied
2. Create complementary probe (antisense), this probe is tagged
3. Add tissue (usually brain slices) and a probe-mRNA duplex forms
Common labels for probes - Answer-Fluorescent dyes, Horseradish peroxidate,
biotin
Immunohistochemistry - Answer-Visualize expression of proteins
1. Incubate sample with primary antibody that recognizes a specific antigen (protein)
2. Perform direct IHC or indirect IHC
Direct IHC - Answer-Labels primary antibody directly
Primates as a Model - Answer-Useful subject to study neuroscience of motor,
sensory, and cognitive tasks (decision making), attention, and learning
Drosophilia as Model - Answer-Test hypotheses involving motor, sensory (vision,
olfaction, taste), memory, and social (courtship and aggression) behaviors
C. Elegans as Model - Answer-Various sensory-motor behaviors can be assessed
(locomotion, mechanical, temperature, and chemical)
Zebrafish as Model - Answer-New, used in developmental studies
General How To Sequence of Animal Surgery - Answer-1. Preoperative procedures
2. Administer anesthesia
3. Perform surgery (from initial incision to last suture)
4. Post-operative care
Preoperative Procedures - Answer-Get animal, prepare surgical area (sterilize)
Anesthesia - Answer-Chemical injection (Ketamine/Xylazine) or gas (Isoflurane)
Perform Surgery - Answer-Place rat in stereotaxic apparatus, usually involves
chronic or acute implant
Post-Operative Care - Answer-Observations, antibiotics
Chronic implants - Answer-Sealable chambers (head wells), cannula, cranial
windows
Acute implant - Answer-Electrolytic cannula
Brain atlas - Answer-3D coordinate system for brain
NEUROSCIENCE EXAM QUESTIONS
AND ANSWERS
Ways to Fix Tissue - Answer-Immersion- place entire specimen in fixative solution
Perfusion- deliver fixative through cardiovascular system
Process of Histology: - Answer-Fixation, embedding, sectioning
Embedding - Answer-Specimens sometimes embedded in gelatin, plastic, par
paraffin wax; stabilizes structure to make it easier to cut
Sectioning - Answer-Cutting of tissue into thin slices using sliding microtome, rotary
microtome, cryostat, vibratome
Sliding microtome - Answer-Frozen, blade moves
Rotary microtome - Answer-Not frozen, in plastic
Cryostat - Answer-Blade is fixed, specimen moves (inverse of microtome), frozen
Vibratome - Answer-Slice fresh tissue, vibrating knife
Why non-fixed tissue? - Answer-If fixative may damage or prevent histological
procedure, can freeze to prevent decay
3 Orientations to cut brain - Answer-Coronal, sagittal, horizontal
Types of Stains: - Answer-Cell body, fiber, golgi
Cresyl Violet - Answer-Cell body stain, stains cell nuclei and nissel, blue to purple,
useful for examining of cytoarchitecture, stains each type of neuron slightly
differently
Hematoxylin - Answer-Cell body stain, stains cell nuclei, blue to blue-black, often
used in combination with Eosin
Eosin Y - Answer-Stains cytoplasm, pink to red
Thionine - Answer-Cell body stain, stains cell nuclei and nissel stain, blue to purple
Weigert - Answer-Fiber stain, stains myelin, normal myeline is blue, degenerated
myelin is yellow, combines with hematoxylin to selectively stain myelin
Golgi stain - Answer-Fills neuron cell bodies and processes, black, stains individual
neurons at random
, Cell body stains- basophilic stains - Answer-Stains acidic molecules like RNA and
DNA (see dendrites, nucleolus, nissel substance, axon hillock, NOT axon
In Situ Hybridization - Answer-Shows where and when a gene is transcribed (mRNA)
1. ID mRNA sequence to be studied
2. Create complementary probe (antisense), this probe is tagged
3. Add tissue (usually brain slices) and a probe-mRNA duplex forms
Common labels for probes - Answer-Fluorescent dyes, Horseradish peroxidate,
biotin
Immunohistochemistry - Answer-Visualize expression of proteins
1. Incubate sample with primary antibody that recognizes a specific antigen (protein)
2. Perform direct IHC or indirect IHC
Direct IHC - Answer-Labels primary antibody directly
Primates as a Model - Answer-Useful subject to study neuroscience of motor,
sensory, and cognitive tasks (decision making), attention, and learning
Drosophilia as Model - Answer-Test hypotheses involving motor, sensory (vision,
olfaction, taste), memory, and social (courtship and aggression) behaviors
C. Elegans as Model - Answer-Various sensory-motor behaviors can be assessed
(locomotion, mechanical, temperature, and chemical)
Zebrafish as Model - Answer-New, used in developmental studies
General How To Sequence of Animal Surgery - Answer-1. Preoperative procedures
2. Administer anesthesia
3. Perform surgery (from initial incision to last suture)
4. Post-operative care
Preoperative Procedures - Answer-Get animal, prepare surgical area (sterilize)
Anesthesia - Answer-Chemical injection (Ketamine/Xylazine) or gas (Isoflurane)
Perform Surgery - Answer-Place rat in stereotaxic apparatus, usually involves
chronic or acute implant
Post-Operative Care - Answer-Observations, antibiotics
Chronic implants - Answer-Sealable chambers (head wells), cannula, cranial
windows
Acute implant - Answer-Electrolytic cannula
Brain atlas - Answer-3D coordinate system for brain
