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Neuroscience > L7 - techniques > Flashcards

L7 - techniques Flashcards

1
Q

electrophysiology

A

measuring electrical activity of biological tissue

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2
Q

potential difference

A

measured between electrode inside and outside of cell

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3
Q

what causes -ve intracellular charge

A

high K+ concentration inside cell

K+ leaves cell via K+ leak channels

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4
Q

3 methods of preparation for measuring electrical changes to neurons

A

1 - dissociated neuron cultures
2 - acute brain slices
3 - whole animal

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5
Q

dissociated neuron cultures method

A

plate fragments of neurons from embryos and add growth medium so they grow into networks

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6
Q

advantages of dissociated neuron cultures

A

easily accessible for recording
cells well isolated
easy to carry out

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7
Q

disadvantages of dissociated neuron cultures

A

only good for embryos

not normal physiological environment for neurons

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8
Q

acute brain slices methods

A

record from neurons in thin slices of brain tissue

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9
Q

advantages of brain slices

A

local circuits remain intact
carry out at any developmental stage
easy to identify specific regions

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10
Q

disadvantages of acute brain slices method

A

long range inputs/outputs get cut

not normal physiological environment

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11
Q

whole animal method

A

animal under anaesthesia
implant electrodes
recordings made when animal is awake

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12
Q

advantages of whole animal recordings

A

all circuits remain intact

allows correlation of electrical activity wth behaviour

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13
Q

disadvantages of whole animal recordings

A

difficult to carry out

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14
Q

2 ways of intracellular recording membrane potential of a neuron

A

sharp electrode

patch clamp recording

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15
Q

sharp electrode method

A

electrode made of glass
filled with solution similar to that of the cell
very high resistance - narrow tip
poke a hole in side of cell
allows access to record membrane potential relative to outside the cell

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16
Q

patch clamp recording

A 
more modern/popular method
lower resistance pipette - wider tip
put on surface of membrane
apply negative pressure (suction)
bursts membrane under pipette
cytoplasm continuous with inside of pipette allows access for recording
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17
Q

equipment required for intracellular recordings

A 
whole set up = rig 
placed on anti-vibration table
microscope
manipulators
big metal cage
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18
Q

how do you isolate pipette from external vibrations

A

rig placed on anti-vibration table

19
Q

what is the microscope for

A

visualising individual neurons within the brain slice

20
Q

what are the manipulators for

A

very delicately place the glass pipette onto the surface of the cell

21
Q

what is the big metal cage for

A

surrounds the rig

isolates rig from ambient electrical noise

22
Q

amplifer

A

takes small analogue/electrical signal and makes it larger

converted to digital signal before being sent to computer

23
Q

why is saline solution used

A

similar to csf

24
Q

cell attached recording

A

pipette attached to membrane via small suction
doesnt break through surface
low resistance seal

25
Q

whole cell recording

A

more suction/negative pressure applied
membrane bursts underneath pipette
allows electrical access to inside of cell

26
Q

inside out patch

A

suction from whole cell recording pulls small bit of membrane from underneath pipette and separates from cell
allows recording of individual ion channels

27
Q

Ohm’s law

A 
V= I x R
voltage = resistance x current
28
Q

low resistance occurs when

A

lots of ion channels are open

29
Q

how do you make current clamp recordings

A

inject different currents to stimulate the cell to give the resistance of the membrane

30
Q

why do you apply different drugs in current clamp recordings

A

drugs alter the ion channels of membranes

causes changes in resistance of membrane

31
Q

describe the EPSP of a CA1 pyramidal neuron

A

fast rising

slow decaying

32
Q

what are voltage clamp techniques for

A

they fix the voltage so it cant change

allows studying of underlying currents

33
Q

the amplifier injects -ve current to maintain cell at -80mV…

A

the cell cant depolarise

opposes +ve ion flow via glutamate receptors

34
Q

EPSC stands for

A

excitatory post synaptic current

35
Q

what is an epsc

A

the flow of ions that causes an epsp

36
Q

as +ve ions move into the cell what happens to the extracellular space

A

becomes more -ve

37
Q

advantages of measuring extracellular space

A

does not disrupt intracellular content
can record spikes in vivo
can record many cells at once

38
Q

what causes the size of the extracellular recording spike to vary

A

the distance of the recording from the electrodes

electrodes pick up spikes closest to them

39
Q

tetrode

A

bundle of 4 electrodes

40
Q

what does functional imaging measure

A

neuronal activity

41
Q

benefits of functional imaging

A

measures a larger anatomical area that electrodes

42
Q

gCaMP

A

calcium indicator protein

fluorescent when bound to Ca2+

43
Q

limitations of gCaMP

A

difficult to distinguish fluorescence between 6 small spikes or 1 large Ca2+ signal

Neuroscience (14 decks)

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