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Center for Microscopy and Image Analysis

Wide-field Nikon-Ti2-E (Irchel)

Nikon Ti2-E

This inverted wildfield system is suited to image live cells as well as fixed samples in fluorescence and transmission light modes. A temperature controlled environment is possible for live cell experiments. The system does not have CO2 control. It is well suited for phase contrast imaging of bacteria and does allow complex imaging tasks.

Responsible Person

responsible person

Location

University Zurich, Irchel Campus, Room Y38-M-77.

Training Request

 Follow this link to apply for an introduction to the microscope

The Nikon Ti2-E  features the following modalities:

Wide-field microscopy

 Fluorescence wide-field microscopy is a technique where the entire specimen is illuminated uniformly, causing fluorophores within the sample to emit light. This emitted fluorescence is then captured by a camera or detector, producing an image that shows the distribution of fluorescent molecules. It's widely used for observing cellular structures, proteins, and other biomolecules tagged with fluorescent markers. The technique offers high sensitivity and allows for real-time imaging of live cells, making it invaluable in biological and medical research.

Differential interference contrast microscopy (DIC)

Differential interference contrast (DIC) microscopy is an advanced optical microscopy technique that enhances the contrast in unstained, transparent specimens. DIC utilizes polarized light split into two beams that pass through the specimen at slightly different angles. When these beams recombine, the differences in optical path length cause interference, creating an image with high contrast and pseudo-3D effect. This technique is particularly useful for observing fine details and structures within live cells and tissues without the need for staining, making it ideal for dynamic studies in cell biology.

Phase contrast microscopy (PH)

 Phase contrast microscopy is a technique designed to enhance the contrast of transparent and colorless specimens, such as living cells, by converting phase shifts in light passing through the specimen into changes in intensity. It employs a phase plate to create constructive and destructive interference, which highlights differences in refractive index within the sample. This allows for the detailed visualization of cellular structures and organelles that are otherwise invisible under standard bright-field illumination. Phase contrast microscopy is widely used in biological research for examining live, unstained cells and tissues.

 

Technical Specifications

Microscope body

  • inverted Ti2-E body

  • Motorized encoded xy-stage and motorized encoded z-drive

  • Perfect Focus System 4

  • Large field of view - 1280 x 1280 pixels, 82um FOV (at 63x)

  • Motorized fluorescence turret, filter-wheel, condenser turret, light path switcher

  • Incubator dark (temperature and humidity control)

Light Sources
  • Halogen lamp for transmitted light
  •  Lumencor SpectraX
Common Fluorophores Excitation LED (nm)
DAPI 395/25 nm 
CFP 440/20 nm 
GFP, Alexa488, Cy2 FITC 470/24 nm 
YFP 510/25 nm 
TRITC, Cy3 550/15 nm
mCherry (on demand) 575/25
DRAQ5, Cy5, Alexa639 640/18 nm

Camera System

  • Orca Fusion CoaXpress sCMOS

  • 16 bit cooled monochrome camera

  • 2304 x 2304 pixel (6.5 x 6.5um pixel size) back-illuminated sCMOS

  • QE: 80 %

  • dynamic range 15 bit

  • Exposure times 17us to 10s (Fast scan)

Environmental control

  • Okolab Cage Incubation with Temperature, and Humidity Control

  • no CO2 control

Accessories
  • Okolab Sample Holders for Wellplates, 35 mm dishes, 1x and 4x Slides

Available Optics
Name Magnification NA Immersion WD (mm)
CFI Plan Apochromat Lambda    2x                            0.1 Air 8.5
CFI Plan Apochromat Lambda DIC  10x 0.45                Air 4.0
CFI Plan Apochromat Lambda PH2  20x 0.75 Air 1.0
CFI Plan Apochromat Lambda DIC  40x 0.95 Air 0.25
CFI Plan Apochromat Lambda PH2  40x 0.6 Air 3.6
CFI Plan Apochromat Lambda PH3 100x 1.45 Oil 0.13

PH = phase contrast, DIC = Differential Interference Contrast

Available Filters

 

MXR00244 4x quad-bandpass: LED-DA/FI/TR/Cy5-4X-B (DAPI / FITC / TRITC / Cy5  - Pinkel Quad)

  
Fluorochromes Reflection to sample (Excitation filter) Transmission to camera (Dichroic)  

DAPI

352-404 nm 

414-450 nm

  

GFP, Alexa488

461-487.5 nm

499.5-530 nm

  
RFP, Alexa568

543-566 nm

580-611 nm

  
Cy5, Alexa647

626-644 nm

 659.5 – 850 nm  

MXR00241 3x triple-bandpass: LED-CFP/YFP/mCherry-3X-A (CFP / YFP / mCherry - Pinkel Triple)
Fluorochromes Reflection to sample (Excitation Filter) Transmission to camera (Dichroic)

DAPI

 426 – 450 nm 464 – 486 nm

GFP, Alexa488

 497.6 – 519.5 nm 532 – 554 nm
RFP, Alexa568 567.4 – 588.6 nm 603 – 800 nm

Emission filters (filters in the filter-wheel) in front of the camera
Position in Filter-wheel Fluorochromes Bandpass Transmission (nm)
1 Transmitted light - empty for 100% transmission
2

DAPI

 single BP  414 – 450 nm
3 GFP, ALEXA 488 single BP 499.5 – 530 nm
4 RFP, ALEXA 568 single BP 580 – 611 nm
5 Cy5, Alexa647 single BP 662.5 - 732.5 nm
6   single BP 626.0 – 644.0 nm
7   single BP 461.0 – 487.5 nm
on request  

Quad-band

414 – 450 nm

499.5 – 530 nm

580 – 611 nm

661 – 800 nm
on request  

Triple-band

464 – 486 nm

532 – 554 nm

603 – 800 nm
on request   single BP 469.5 – 494.5 nm
on request   single BP 532 – 556 nm
on request   single BP 604 – 679 nm

 

User Guide

 will come soon!

Links & Literature

Nikon Ti2-E
 

Sample preparation
 

Filter example
 

Spectra viewer

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