Widefield inverted

Introduction

Two inverted systems for transmitted and fluorescence widefield microscopy of fixed specimens and short or extended time lapse of live samples.

(1) Time lapse 1

Hardware specifications

The Zeiss Axio observer microscope in Time lapse 1 system is wholly enclosed in a polycarbonate incubator system with controlled temperature and humidity and an active gas mixer. It features a motorized stage, X-cite metal halide light source and a monochrome AxioCam HRm

 

Reflector Turret Excitation Beamsplitter Emission Suitable fluorophores
Analyzer module DIC
Zeiss Filter set 50 (optional) BP 640/30 FT 660 BP 690/50 Cy5, DRAQ5
Zeiss Filter 20HE BP 546/12 FT 560 BP 607/80 PI, Cy3, Alexa555, Rhodamine
Zeiss Filter set 38HE BP 470/40 FT 495 BP 525/50 Cy2, GFP, Alexa 488
Zeiss Filter set 49 G 365 FT 395 BP 445/50 DAPI, Hoechst
Optovar x1.6

 

Objective Magnification NA Contrast Dish bottom or slide coverslip Working distance (mm) Immersion
EC Plan-Neofluar x5 0.16 Phase 1 plastic/glass 18.5 air
EC Plan-Neofluar x10 0.3 Phase 1 plastic/glass 0.3 air
Plan-Neofluar x20 0.4 Phase 2 plastic/glass 0.4 air
Apochromat x20 0.8 Phase 2 & DICII Glass 0.55 air
Apochromat x40 0.95 Phase 3 Glass 0.25 air
Plan-Apochromat (optional) x63 1.4 Phase 3 Glass 0.19 oil

 

We have glass bottom dishes for you to try imaging with the higher end objectives 

Acquisition

Acquisition is performed with the Zeiss proprietary Axiovision software. Output files can be opened by the ZEN lite edition software, ImageJ/FIJI or exported to TIFF for analysis.

Special applications

Long term live experiments in 6 dimensions: XYZ, positions, channels, time, in optimal gas composition or hypoxia. Also FRET, FRAP, wound healing, tube formation. 

(2) Time lapse 2

The Zeiss Axio observer inverted microscope system is suited for fixed specimens and for short term live experiments on 35mm dishes using a stage mini incubator. The system features a motorized stage, X-cite metal halide light source and Colibri LED illumination, high resolution monochrome Hamamatsu Orca R2 camera , color Axiocam MRc 5 camera  and high speed Axiocam HS. 

Reflector Turret Excitation Beamsplitter Emission Suitable fluorophores
Zeiss Filter set 49 G 365 FT 395 BP 445/50 DAPI, Hoechst
Zeiss Filter set 45 BP 560/40 FT 585 BP 630/75 Mitotracker red, mCherry, Texas Red
Chroma 49005 ET 545/30 T 570 LP ET 620/60 Cy3, Mitotracker Orange, DsRed
Zeiss Filter 20HE BP 546/12 FT 560 BP 607/80 PI, Cy3, Alexa555, Rhodamine
Zeiss Filter set 38HE BP 470/40 FT 495 BP 525/50 Cy2, GFP, Alexa 488
Analyzer module DIC
Optovar x1.6
Zeiss Filter set 47 (optional) BP 436/20 FT 455 BP 480/40 CFP
Zeiss Filter set 61HE
(Colibri) (optional)
BP 474/28@LED470
BP 585/35@LED590
DFT 495+605 DBP 527/54+645/60 GFP/mCherry FRET
Zeiss Filter Set 48 (optional) BP 436/20 FT 455 BP 535/30 CFP/YFP FRET
Zeiss Filter Set 59HE (optional) BP 445/25@LED455
BP 510/15@LED505
DFT 460+520 DBP 480/22+LP530 Dual CFP/YFP

 

Objective Magnification NA Contrast Dish bottom or slide coverslip Working distance (mm) Immersion
EC Plan-Neofluar x5 0.16 Phase 1 plastic/glass 18.5 air
EC Plan-Neofluar x10 0.3 Phase 1 plastic/glass 0.3 air
Plan-Apochromat x20 0.8 DICII Glass 0.55 air
EC Plan-Neofluar x40 0.75 DICII Glass 0.71 air
Plan-Apochromat x63 1.4 Phase 3 Glass 0.19 oil
EC Plan Neofluar x100 1.4 BF Glass 0.17 oil

 

We have glass bottom dishes for you to try imaging with the higher end objectives 

Special applications

Short term live imaging with heated stage insert with humidity and fixed gas inlet. Tiling, positions, Z-stacks, photo activation, FRET, high speed imaging (up to 60 f/s full frame), color brightfield.

Acquisition

The Zen 2 software provides fast acquisition, tiling and positions, multidimensional and fast acquisition and basic 2D analysis.

Analysis 

For basic visualization and processing you can download the free ZEN lite edition software. Output files can be opened by ImageJ/FIJI and Imaris 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.

For reference you may use the Time Lapse 1 instruction file  or the Time Lapse 2 instruction file.

Reservations

For long term live widefield microscopy, Time Lapse 1 can be reserved in the BookItLab online reservation system, Microscope Time-lapse 1.

For general widefield microscopy and for tiling, color brightfield, fast acquisition, physiology, Time Lapse 2 can be reserved in the BookItLab online reservation system, Microscope Time-lapse 2. For live experiments, the heating stage insert should be reserved separately.

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