Confocal LSM 700

 

Introduction

In confocal microscopy, placement of a variable spatial pinhole at the confocal plane of the lens eliminates out of focus light thus reducing background interference.

The Zeiss LSM700 is a laser scanning confocal system attached to an upright motorized microscope (Axio Imager.Z2). Its innovative variable secondary dichroic beamsplitter splits emission signals between the two PMTs and the cutoff can be freely selected in 1nm steps.

 

Laser line Wavelength Power Suitable fluorophores
Solid state laser cassette 633 nm 5mW Cy5, APC, Alexa 647, TO-PRO-3
Solid state laser cassette 555 nm 10mW Cy3, TAMRA, PI, Texas Red, PKH26
Solid state laser cassette 488 nm 10mW Cy2, GFP, Mitotracker Green, JC1
Solid state laser cassette 405 nm 5mW DAPI, Hoechst

 

Objective lens Magnification NA coverslip Working distance Immersion
EC Plan Neofluar x10 0.3 0.17 mm 5.2 mm Air
Plan Apochromat x20 0.8 0.17 mm 0.55 mm Air
LD LCI Plan Apochromat x25 0.8 0-0.17 mm 0.57 mm Oil Glycerin Water
EC Plan Neofluar x40 1.3 0.17 mm 0.21 mm Oil
W Plan Apochromat x40 1.0 2.5 mm Water dipping
Plan Apochromat x63 1.4 0.17 mm 0.19 mm Oil
C Apochromat (optional) X40 1.2 0.14-0.19 0.28 mm Water

Acquisition

Acquisition is performed with the Zeiss proprietary ZEN 2010 software.
Two PMT detectors for simultaneous detection and up to four sequential tracks and one transmitted-light PMT allow superimposing fluorescence images on brightfield or DIC images.
The software allows 360° XY scanning field rotation, and freely definable regions of interest (ROI).
Acquisition modes: Spot, line/spline, frame, lambda stack.
Resolution up to 2048 x 2048 pixels, scanning speed up to 5 frames/sec at 512 x 512 pixels.

Special applications

Photobleaching, live imaging, ROI, positions, tiling, time series and combination (XYZ|T), increased intensity over Z.

Analysis 

The Zen software provides a wide range of 2D/3D image processing tools: export to TIFF, maximum projection, 3D reconstruction, 2D colocalization, spectral unmixing and more. For basic visualization and processing you can download the free ZEN lite edition software. An offline full version is available for analysis at Computer Analysis 4. Output files can be opened by Imaris and ImageJ/FIJI for analysis.

Instructions for specific applications

New users

Please contact Edith Suss-Toby, tel. +972-4-8295347/61 to coordinate a meeting.
Tips for immunofluorescence experiments.
Please prepare controls before starting a new model.

Reading materials prior to the first instruction session:

Reservations

The system can be reserved in the BookItLab online reservation system, Confocal LSM 700  upright microscope. For live experiments, the heating stage insert should be reserved separately.

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