This Zeiss LSM700 confocal is mounted on an inverted Zeiss Axio Observer Z1 using a motorized XY stage controller. We have 4 solid state laser lines (445, 488, 555 and 639nm) available on this system.
Currently four objectives are on the system: a Plan Apo 10x Ph1 objective with a 0.45 NA, a Plan Apo 25x Oil immersion DIC objective with 0.8 NA, a Plan Neofluar 40x Oil immersion DIC objective with a 1.3 NA and an Apochromat 63x Water immersion DIC objective with a 1.2 NA.
Two photomultiplier detectors are available, each coupled with 8 emission filters for spectral flexibility on the detection of the fluorescence signal. An additional transmitted light detector may also be selected to allow the users to acquire a transmitted light image of the samples in confocal mode.
The software running the LSM700 is the ZEN ("Zeiss Efficient Navigation") software. That software allows a really easy and straightforward use of the confocal. Using the so-called "Smart Setup" function, the software will drive you in a few clicks through the light path configuration to set up for your own experiments.
Additionally, the focus stabilizer ("Definite Focus") implemented on this confocal microscope enables maintaining the focus stable in the z axis for in long term (typically time-lapse) acquisitions.
Confocal Microscope Leica SP 8 FLIM
This confocal microscope comes with a pulsed
supercontinuum (White Laser Line) tunable laser in the visible range from 470nm
to 670nm. Tune freely your own BP filter settings for detection that are
matching for your fluorescent detection. The fine tunable WLL (every wavelength
with 1nm accuracy can be selected from the 470-670nm range) will allow to
perform any type of excitation within this range and also measuring the
excitation spectrum of your fluorescent dyes. Complete excitation emission
spectra can be recorded.
A FLIM (Fluorescence Lifetime Imaging) module
is also attached to this system. So with that you can measure precisely
fluorescence lifetime of your dyes and perform Single Molecule Detection (SMD).
This feature is fully integrated to the confocal system (developed by PicoQuant
company using Symphotime software). This SMD FLIM can also be used to make FRET
experiments.
The microscope stand is an inverted one fully
motorized. The system has a motorized xy stage controller. The motorized z
focus can use the microscope motorization or the table top piezo z for fine and
fast z movements.
On the detection side of the fluorescence of
your samples its fully filter free you can design and fine tune your emission
spectrum window freely. Two sensitive HyD + 1 HyD-SMD/FLIM detectors (Hybrid Detectors)
are installed with one standard PMT detector. The HyDs are special detection
devices that will allow you to time shift the detection of the light coming
from the sample using the so called gating feature (adjusting in time gate
switches off the data collection during the WLL laser pulse) this will help in
many cases to get read of autofluorescence/biological background of the sample
used. The HyDs are also generating no electronical noise and can be used for
photon counting experiments.
A transmitted light PMT is added to the system
for the confocal brightfield recordings.
As complementary lasers on the Leica SP8 we
have an Argon ML laser generating the 458, 488 and 514nm lines. A 440nm solid
state laser is also present for the FLIM measures.
On the objectives side the system is equipped
with a:
10x
HC PL FLUOTAR NA 0.30 Dry,
20x
HC PL NA 0.75 IMM,
40x
HC PL IRAPO NA 1.10 WATER,
63x
HC PL APO NA 1.20 WATER.
In summary this system will enable you to
perform all the traditional confocal multi-dimensional experiments as
consecutive optical slices (z-stacks), xy tiles stitched automatically
together, time lapses, fluorescent multi channels acquisitions, spectral
characterized experiments and all these can be combined. Also precise FRAP/FRET
or FLIM experiments can be performed.
This brand new system is now installed at the CIF
Dorigny campus (Biophore, Room 4416).
MICROSCOPES - GENOPODE
Confocal Microscope Zeiss LSM 510 Meta INVERTED
Available Laser lines :
Diode 30mW 405 nm
Argon Multiline 30mW 458/477/488/514 nm
HeNe 1mW 543 nm
HeNe 2mW 594 nm
HeNe 5mW 633 nm
Equiped with transmited detector for DIC and Phase Contrast.
Meta detector for emission spectra of fluorochromes.
Available visible laser lines 458, 488, 514, 561 and 633 nm
for conventional laser scanning confocal microscopy.
The available laser for the 2 Photon acquisitions is a Ti:Sapphire
Chameleon Ultra II infrared laser, tunable from 690 nm to 1040 nm with an maximum output
power of 4W at 800 nm.
Microscope
An upright Zeiss AxioExaminer Z1.
Objectives available on the system, currently 3 objectives
are available on the system:
Achroplan 10x W objective with a 0.3 NA
Plan-Apochromat 20x W objective with a 1.0 NA
Apochromat 63x Water-Oil immersion objective with a 1.2 NA
Attached Software
ZEN 2009 (Zeiss Efficient Navigation 2009)
Detection modes
The light may be detected in the confocal scanhead (internal
or 'descanned' detection)
Confocal detection on external 'non-descanned' detectors
(NDD)
Confocal detection on GaAsP non-descanned detector (GaAsP
NDD)
System Capabilities
In depth confocal microscopy for thick samples 3D optical
sectioning.
Spectral infrared excitation fingerprinting.
Spectral emission fingerprinting.
FRAP, FRET, Time lapse, fluorescent dyes uncaging
Confocal Microscope Zeiss LSM 510 Meta UPRIGHT
Available Laser lines :
Diode 30mW 405 nm
Argon Multiline 30mW 458/477/488/514 nm
HeNe 1mW 543 nm
HeNe 2mW 594 nm
HeNe 5mW 633 nm
Equiped with transmited detector for DIC and Phase Contrast.
Meta detector for emission spectra of fluorochromes.
Attached software : LSM Software Rel. 3.5
UPRIGHT Microscope
AXIO Imager Z.1
Fully motorized
Filter cubes
Set 49 DAPI Ex G 365, FT 395, EM BP 445/50
Set 38 HE eGFP EX BP 470/40, FT 495, EM BP 525/50
Set 43 HE Cy3 Ex BP 550/25, FT 570, EM BP 605/70
Set GFP Chroma
Objectives
EC Plan-NEOFLUAR 10x/0,30
ACHROPLAN 10x/0,30 W Ph1
ACHROPLAN 20x/0,50 W Ph2
ACHROPLAN 40x/0.80 W
EC Plan-NEOFLUAR 40x/1,30 Oil DIC
ACHROPLAN 63x/0,95 W
Plan-APOCHROMAT 63x/1,40 Oil DIC
Camera : AxioCam MRm Axiovision Software Rel. 4.4
Illumination: HAL 100, HBO 100
Stereomicroscope Leica MZ16FA
Illumination Standard + Fluorescence Fluorescence Leica EL6000 (lasts up to 2000 hours)
Magnification
1x up to 180x
Filters
DAPI, GFP, CY3, Transmitted light
Attached software : Leica Application Suite (LAS)
Cameras :
Leica DFC350FX Monochrome
Leica DFC320R2 Color
Motorized filter wheel, focus and zoom.
Laser Microdissection and laser tweezers
Microscope :
Inverted OLYMPUS IX71
Illumination:
HBO,
HAL
Filters cubes for Dapi, FITC, TRITC:
narrow-band DAPI Ex BP360-370, BS DM400, Em BA420-460
wide-band FITC Ex BP460-495, BS DM505, Em BA510-550
wide-band TRITC Ex BP530-550, BS DM570, Em BA575-625
Objectives:
Plan Fluorite 4X/0.13 PHL wd 17.00mm
Plan Fluorite 10X/0.30 PH1 wd 1.60mm
Plan Fluorite 20X/0.45 PH1 wd 6.40 – 7.6mm.60mm
Plan Fluorite 40X/0.60 PH2 wd 2.70mm – 4.00mm
Plan Fluorite 60X/0.70 PH2 wd 1.50mm – 2.20mm
Plan Super Apochromat 100X/1.40 oil imm wd 0.12mm
Modules :
mmi CellCut® Laser Microdissection System
Computer controlled xy-table
mmi CellManipulator Laser tweezer
Multi Cap System
Multi-Slide System
Attached Camera :
3 Chip RGB CCD Sony camera
20´´ Interactive Pen-Screen Display
Laser for cutting and isolation of tissue samples :
Solid state UV-A laser, 350 nm
NIH Image / Scion Image / Visilog / Image J
General purpose image analysis software with programming language.
Bitplane IMARIS suite 4.1
Consisting of Imaris, ImarisMeasurement Pro, ImarisTrack and ImarisColoc.
Large range of specific image file readers for 3D microscopes. SLicer, Section, Gallery,
Easy 3D shadow Projection, real time volume rendering.
Interactive 3D distance measurement, volume and voxel intensity statistics in 3D and 4D,
manual object segmentation with contours. Leading 3D and 4D colocalization.
Bitplane ImarisXT - programing interface Bitplane Filament Tracer 4.1, to segment and visualize filamentous structures AutoAligner 2.0, for automatic and manual registration of serial 2D images
SVI HUYGENS Essential Rel. 2.8
3D Deconvolution software
Huygens® Essential is an image processing software package tailored for deconvolution of microscopic images.
Huygens Essential is able to deconvolve a wide variety of images ranging from 2D widefield (WF) images
to 4D multi-channel two-photon confocal images. To facilitate comparison of raw and deconvolved data,
or results from different deconvolution runs the Essential is equipped with a dual 4D slicer tool.
Powerful and fast renderers produce different representations of your data, not needing any special graphic card.
Xenogen Living Image 2.5.1
Xenogen 3D analysis package
Imaging software from the Whole animal imaging system, IVIS 3D from Xenogen.
