Microscopy

Stereomicroscope

This microscope is better for bigger, bulkier objects which means it has a large working distance but lower resolution.

6.3-40x

What is the zoom range for a stereomicroscope?

8-100x

What is the magnification for a stereomicroscope?

Transmitted light

Which light is below going up?

Reflected light

Which light is up going down?

Oblique light

Which light comes in at a different angle?

eyepiece x objective

How do you determine the magnification of a microscope?

To compound light

Why does a compound microscope have multiple objectives?

Objectives are facing down

What does it mean when a compound microscope is upright?

Low

Big working distance = ___ resolution

High

High magnification = ____ resolution

40-1000x

What is the magnification of a compound microscope?

2D

Is a compound microscope 3D or 2D?

Glass transmits light, plastic distorts

Why are glass slides better than plastic slides?

Resolution

What gives us the ability to distinguish between two points?

Numerical aperture (NA)

What is the light gathering ability that is directly correlated with resolution?

Depth of field

What depth is focused on volume of the objective (volume in focus)?

Depth of focus

What depth is the 0 point, above and below (where the center point is the depth of field)?

Field of view

What is the volume visible from side to side and up & down?

Contract

Black vs white

Working distance

What is the volume from the top of the sample to the bottom of the objective?

Line pair

What is the common way to test for resolution?

Empty magnification

What is it called when the object is over-magnified and hazy (pushing resolution beyond the max)?

Par focal

What is it called when focus remains the same through all objectives?

Par central

What is it called when the objectives remain centered?

Field irus

What limits light by opening and closing (field of view to open & close). ITS HOW YOU KOHLER A MICROSCOPE

Condensor irus

What changes contrast?

Condensor

What aligns light?

Kohler illumination

What is the alignment of light from the condenser to the objective? It optimizes light on every plane

Use 10 objective and ensure sample is in focus

What is step 1 of kohlering a microscope?

Close field irus

What is step 2 of kohlering a microscope?

Center and focus the condenser

What is step 3 & 4 of kohlering a microscope?

Reticle

What is the focus of the eyepiece?

Your eye, the sample, and the field irus

What 3 things are in the visible plane?

near focal plane of objective, condenser iris, and light bulb/source

What 3 things are in the nonvisible plane? (when you remove one eyepiece)

Diffuser

What part evens out (diffuses) light by spreading evenly?

0.99

What is the maximum numerical aperture achieved with a dry objective?

Immersion method (oil)

Any numerical aperture above 1 requires what?

The condenser focuses and directs light, numerical aperture determines how much light the microscope can collect and resolve.

What is the relationship between numerical aperture and condenser?

Brightfield

Which contrast method is the most common, requires no external components, and floods the sample with light? (resolution = high)

Darkfield

Which contrast method uses oblique lighting which causes everything to go black & white? It requires a condenser slide (non-visible plane) and resolution is low.

Phase contrast

Which contrast method requires a phase slider and phase objective? It has a blue background and can help increase contrast in translucent samples. Resolution = low. OBTACLE STAINING

Polarization

Which contrast method eliminates certain direction of light (orients light)? It presents colors or an extinction point. It requires an analyzer and polarizer. Resolution = mid

Differential Interference Contrast

Which contrast method adds dimension to a non-dimensional sample? It requires an analyzer, polarizer, condenser prism, and an objective prism. Resolution = high

Fluorescence

Which contrast method cuts out the condenser and uses objective x2 to eliminate everything but a small amount of light? It requires a white light source, a fluorescence filter, and sometimes fluorescence dye. Resolution = highest

Brightfield, none

Which contrast method is this? What external components does it require?

Darkfield, condenser slide

Which contrast method is this? What external components does it require?

Phase contrast, phase slider/phase objective

Which contrast method is this? What external components does it require?

Polarization, analyzer/polarizer

Which contrast method is this? What external components does it require?

Differential interference contrast, analyzer/polarizer/condenser prism/objective prism

Which contrast method is this? What external components does it require?

Fluorescence, white light source/fluorescence filter, fluorescent dye

Which contrast method is this? What external components does it require?