Introduction; Hardware; Objectives; Filter Turret - Zeiss LSM 700 Instructions Manual

Introduction
The Zeiss LSM 700 is equipped with 4 laser lines and can detect up to four color signals at frame rates
approaching 5 frames per second at 512 x 512 pixels. Efficient separation of the fluorescence signals by
selective laser excitation coupled to efficient splitting of the emission using the variable secondary
dichroic (VSD) beamsplitter prevents crosstalk and enables spectral imaging as well as linear unmixing of
highly overlapping fluorophores. Among the advanced features of the VSD beamsplitter is that all portions
of the emission spectrum are utilized for determining each spectral data point.
Hardware
The microscope:
The LSM 700 is attached to an upright microscope (Axio Imager Z2).
Objectives
Position Objective lens
1 W Plan
Apochromat
2 EC Plan
Neofluar
3 Plan
Apochromat
4
LD LCI Plan‐
Apochromat
5 EC Plan‐
Neofluar
6 Plan
Apochromat
Filter turret
position Filter set
1 FS38
2 FS43
4 Fs49
5 DIC
Magnification NA coverslip
X40 1.0
x10 0.3 0.17 mm
x20 0.8 0.17 mm
x25 0.8 0‐0.17
x40 1.3 0.17
x63 1.4 0.17
Excitation filter Beam splitter
BP 470/40
BP 545/25
G 365
Working
distance
0 2.5 mm Water dipping
5.2 mm
0.55 mm
0.57 at cover
Glycerine and
glass 0.17
0.21mm
0.19 mm
Emission filter
FT 495 BP 525/50
FT 570 BP 605/70
FT 395 BP 445/50
Parfocal
Immersion
length
45 mm
45 mm
Air
45 mm
Air
Water, 45 mm
Oil
45 mm
Oil
45 mm
Oil
Fluorophore
family
Cy2,GFP
Cy3,Al555
DAPI,BFP
p. 2