Microscopy Flashcards

1
Q

Microscope

A

An instrument that allows you to magnify an object that may not be visible with the naked eye.

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2
Q

Types of Microscopes (4)

A

Light
SEM
TEM
LSCM

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3
Q

SEM

TEM

A

Scanning Electron Microscope

Transmission Electron Microscope

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4
Q

Eyepiece Lens

A

x10 Magnification

Eyepiece graticule can be inserted

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5
Q

Coarse-focus knob

A

Allows for quick focus

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6
Q

Fine-focus knob

A

Allows for intricacy in focusing on an image

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7
Q

Turret

A

Allows to switch between different objective lenses

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8
Q

Objective Lens

A

x4,x10,x40

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9
Q

Specimen Stage

A

Where slide is kept

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10
Q

How a microscope is held

A

Arm and Base

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11
Q

How an SEM works

A

A beam of electrons sent across the surface of a specimen.

Reflected electrons collected

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12
Q

How a TEM works

A

A beam of electrons is transmitted through a specimen and focused to produce an image.

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13
Q

How an LSCM works

A

A single spot of focused light is moved across a specimen, causing fluorescence from components labelled with a dye.
Emitted light from specimen filtered through a pinhole aperture.
Only light radiated from near to the focal plane is detected.

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14
Q

Properties of a Light Microscope

A

Wavelength is longer
Resolution of 200nm
Max Magnification of x2000
Chemical Stain

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15
Q

Properties of a TEM

A

shorter wavelength
Resolution of 0.2nm
Max Magnification of x2,000,000
Heavy Metal Stain

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16
Q

Properties of an SEM

A

the same wavelength of TEM
Resolution of 3nm
Max Magnification of x500,000
Heavy Metal Stain

17
Q

Properties of LSCM

A

Uses Light

Allows for 3D imaging whilst object can be alive

18
Q

Light Microscope Image

A

2D
Organelles
Transparent until Stained
The specimen can be alive

19
Q

TEM image

A

2D
Organelles
Black and White unless stained
The specimen is not alive

20
Q

SEM image

A

3D
No Organelles: only Surface
Black and White unless stained
The specimen is not alive

21
Q

LSCM

A

3D
produces z sections
Colored with ‘dye’
The specimen can be alive

22
Q

Properties of Staining

A

Increases contrast within cells

Allows components to become visible

23
Q

Differential Staining

A

Used to distinguish between two types of organisms that would otherwise be hard to identify.

24
Q

Gram stain

A

Crystal violet and then iodine is applied to the slide.
Gram-positive retain the stain- purple
Gram negative do not- too thin of a cell wall- Sarafin (stain) turns pink

25
Acid Fast acts on
Differentiates Species of Mycobacterium from other bacteria
26
Acid Fast Process
Lipid solvent is used to carry carbolfuchsin dye. Cells washed with a dilute acid-alcohol solution. Mycobacterium not affected by wash and retain stain.
27
Magnification=
Size of image/Actual size
28
Actual Size=
Size of Image/Magnification
29
Size of Image=
Magnification x actual size