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DMI4000 Microscope

The SPE confocal is built around a motorised DM4000 microscope stand. The fluorescence filter turret and objective lens nosepiece are manually rotated. There is currently no internal focus motor; confocal sectioning is achieved via a galvanometer driven stage insert. Eyepiece or confocal light path selection is automated. DIC is available on this microscope.

Objective Lenses on the Leica DM4000 Microscope

Leica ACS objective lenses have been designed specifically for SPE confocal microscopes with 405 nm lasers to correct for colour registration errors between the violet and visible lines. They do not have a flat field so Leica recommends that the confocal zoom is set to ≥1.6.

Magnification

Immersion Medium

Designation

Numerical Aperture

Coverslip Correction

DIC Prism

10X

Air

ACS APO

0.3

None

C

20X

Multi Imm

ACS APO

0.6

0 - 0.17


40X

Oil

ACS APO

1.15

0.17

E

63X

Oil

ACS APO

1.3

0.17

E

Fluorescence Filter Sets on the Leica DMI4000 Microscope

Turret Position

Filter Set

Excitation Filter

Dichroic Mirror

Emission Filter

Example Fluorophores

1

A

BP340-380

RKP400

LP425

Hoechst/DAPI

2

I3

BP450-490

RKP510

LP515

FITC/GFP

3

N2.1

BP515-560

RKP580

LP590

Texas Red

4

BF

N/A

N/A

N/A

Transmitted Light

5

N/A

N/A

N/A

N/A

Empty

6

Pol

N/A

N/A

N/A

Polariser for DIC and polarised light microscopy

Positions 4 to 6 are empty and not used in this stand.

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