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Leica
Location: MRC Building room 3.17
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This
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is
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a
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Leica
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DMIRB
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inverted
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research
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fluorescence
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microscope
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suitable
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for
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observing
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cells
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on
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slides
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or
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in
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glass-bottomed
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or
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plastic
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bottomed
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vessels
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using
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either
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phase
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contrast
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or
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fluorescence
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microscopy.
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The
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microscope
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has
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a
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Hamamatsu
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C4742-95
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Orca
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greyscale
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camera
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suitable
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for
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acquiring
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images
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of
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fluorescent
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specimens
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but
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not
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suitable
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for
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imaging
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histologically
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or
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histochemically
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stained
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specimens.
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The
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microscope
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has
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20x
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and
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40x
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long
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working
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distance
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objective
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lenses
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that
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have
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adjustable
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correction
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collars
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so
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the
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objectives
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can
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be
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corrected
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to
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focus
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through
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different
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thicknesses.
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For
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example,
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coverslips
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are
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usually
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only
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170
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microns
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thick
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but
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the
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growth
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surface
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of
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standard
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cell
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culture
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dishes
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is
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approximately
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1
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mm
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thick,
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so
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a
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correction
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collar
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would
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have
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to
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be
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set
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differently
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for
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specimens
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mounted
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on
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one
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or
...
other
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of
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these
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surfaces.
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| Table of Contents |
|---|
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|
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|
Objective lenses
Magnification | Immersion | Designation | Contrast Technique | Coverslip Correction (mm) | WD (mm) | |
|---|---|---|---|---|---|---|
10x | Air | N Plan | 0.25 | Ph1 | None | 17.6 |
20x | Air | N Plan LWD | 0.4 | Ph1 | 0 - 2 | 3.2 - 1.9 |
40x | Air | N Plan LWD | 0.55 | Ph2 | 0 - 2 | 3.3 - 1.9 |
20x | Air | N Plan LWD | 0.4 | LMC | 0 - 2 | 3.2 - 1.9 |
40x | Air | N Plan LWD | 0.55 | LMC | 0 - 2 | 3.3 - 1.9 |
Fluorescence filter sets
Filter Position | Filter Set | Excitation Filter | Dichroic Mirror | Emission Filter | Fluorophores |
|---|---|---|---|---|---|
1 | Leica A4 | 360/40 | 400 | 470/40 | DAPI, Hoechst |
2 | Leica GFP | 470/40 | 500 | 525/50 | FITC, GFP |
3 | Leica N2.1 | 515 - 560 | 580 | LP 590 | Cy3, TRITC |
4 | Blank | N/A | N/A | N/A | Transmitted Light |
Camera Calibrations
Magnification | 100 pixels = | 1 pixel = | |
|---|---|---|---|
10x | 1 | 106.34 µm | 1063.4 nm |
20x | 1 | 53.17 µm | 531.7 nm |
40x | 1 | 26.59 µm | 265.9 nm |
10x | 2 | 212.68 µm | 2126.8 nm |
20x | 2 | 106.34 µm | 1063.4 nm |
40x | 2 | 53.17 µm | 531.7 nm |