Nikon microscope 4

Location: HWB room 3/51
Contact: Dr Kees Straatman (krs5).

A free NIS-Elements viewer can be downloaded from here

With recent funding from the University of Leicester this microscope system has been upgraded at the beginning of 2016 with a PFS hardware auto-focus system + software upgrades, new objectives, a Hamamatsu ORCA-Flash 4.0v2 sCMOS camera and a new computer system.

This is a Nikon Eclipse Ti-E microscope stand equipped with COOLLED PrecisExcite High Power LED Fluorescent Excitation System. LEDs are very stable, long lasting, good for live cell imaging and the microscope can be used as soon as the LEDs are switched on. Three wavelengths are available: 400nm, 490nm and 565nm. LEDs with other wavelengths are available and can be added to the system; see CoolLED.

There are sample holders available for slides, 35 mm dishes and multi well plates.

The microscope has a motorized piezo stage for fast Z acquisition with encoders for accurate multiposition imaging. After the upgrade the system has two cameras. At the left port is an Andor iXon EM-DU897 camera, 512 x 512 pixels (16 µm x 16 µm) and a quantum efficiency of > 90%, at the right port the new Hamamatsu ORCA Flash 4.0v2 sCMOS camera. This camera has a very large field of view of  2048 × 2048 pixels (6.5 μm×6.5 μm) and a quantum efficiency peak of 82%.

Super resolution reconstruction

Nikon4 can also be used to acquire images for super resolution reconstruction using Super-Resolution Radial Fluctuations calculations using the ORCA Flash 4 camera. This is possible for both fixed and live samples using any of the colours you would normally use for this system. Please see this page.


Available objectives:
CFI Plan Fluor 4х/0.13 - WD = 17.2 mm
CFI Plan Fluor DLL 10x/0.13 Ph1 - WD = 16 mm
CFI S Plan Fluor ELWD 20хC/0.45 - WD = 6.9 - 8.2 mm
CFI Plan Fluor ELWD 40x/0.6 Ph2 - WD = 2.8 - 3.6 mm
CFI Plan Apo VC 60x oil/1.4 DIC N2 - WD = 0.13 mm
CFI Plan Apo VC 100x oil/1.4 DIC N2 - WD = 0.13 mm

Filter cubes available on this system:

Position1: V-2A filter cube: DAPI
Excitation filter N EX380-420
Dichroic mirror N DM430
Barrier Filter N BA450

Position 2: 31044v2 : CFP
Exciter D436/20x (426 - 446 nm)
Dichroic 455DCLP
Emitter D480/40m (460 - 500 nm)

Position 3: 11001v2 filter cube : FITC/GFP
Exciter D470/40x (450 - 490 nm)
Dichroic 495DCLP
Emitter E515LPv2 (Long Pass filter! Band Pass filter at Position 6!)

Position 4: 41043 HcRed1 - RFP/Red
Exciter HQ575/50x (550 - 600 nm)
Dichroic Q610LP
Emitter HQ640/50m (615 - 665 nm)

Position 5: 49008- ET mCherry, Texas Red
Exciter ET560/40x (540 - 580 nm)
Dichroic T585lpxr
Emitter ET630/75m (592 - 667 nm)

Position 6: 49002- ET GFP- FITC/Cy2
Exciter ET470/40x (450 - 490 nm)
Dichroic T495lpxr
Emitter ET520/50m (500 - 550 nm) (Band Pass filter! Long Pass filter at Position 3)

For live cell imaging an environmental chamber with temperature and CO2 control is included. The microscope is controlled via the Nikon NIS-elements software.


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