Widefield microscopy is a commonly used method for viewing dynamic and topographical information which encompasses many different specialities including brightfield, differential interference contrast (DIC), phase-contrast and epifluorescence. In a widefield microscope, the entire sample is illuminated and the sample is optically sectioned purely via the objective lens. These systems are typically faster than confocal; however, samples are hit with more light and often have a lower signal: noise ratio.

Most widefield microscopes have two light sources, a halogen bulb for brightfield, and a much brighter fluorescence light source (usually a mercury lamp). In the example upright widefield microscope, the brightfield lamp is housed in the base of the microscope and passes through a field stop and condenser before hitting the sample. In comparison, the fluorescence light source is at the rear of the microscope and passes through a filter cube before being focused by the objective onto the sample.

In fluorescence microscopy, light of a particular wavelength is used to excite a molecule which in turn emits light at a different wavelength before being detected via a camera. Filter cubes contain two glass filters and a special mirror called a di-chroic. Unfiltered (white) light first enters the cube via an excitation filter (this only allows light from a narrow range of the spectrum through). The di-chroic mirror reflects particular wavelengths of light, while allowing others to pass through. Here the green excitation light is reflected, sending it down to the sample. Emission light (longer wavelength than excitation light) that comes back from the sample can pass freely through the di-chroic and is then filtered with the emission filter. Typically our microscopes hold 4-6 filter cubes often suited for a wide variety of dyes and fluorophore combinations such as DAPI, CFP, GFP/Alexa488, YFP, RFP/mCherry, or Cy5/Alexa647.

BrightField/DarkField (BF/DF)

Current Status:  Fully Operational


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Room 4.022 - Olympus BX51 Upright Microscope Stand with Olympus DP70 CCD camera

Upright widefield microscope with Halogen lightsource and CCD camera. Suited for imaging fixed samples on slides.

Features Include:

  • Manual X-Y-Z stage

Transmitted Light:  Olympus 100W Halogen Lamp

Condensor:

Name N.A. W.D. mm
Brightfield 0.90 -

Objectives:

Position Objective Magnification N.A. Immersion W.D. (mm) Type Type Thread
1

PlanN

2x

0.06

Dry

-

-

 

M27

2

UPlanApo

4x

0.16

Dry

-

-

 

M27

3

PlanN

10x

0.25

Dry

-

-

 

M27

4

UplanSApo

20x

0.75

Dry

-

-

 

M27

5

UPlanSApo

40x

0.90

Dry

-

-

 

M27

6

-

-

-

-

-

-

 

M27

Computer:

Dell PC - 3GHz Xeon Processor - 8GB RAM - 256GB nVidia Quadro GPU

1TB Raid Array

30" LCD Monitor

Software: 

DP Manager

 

Fluoro 1

Current Status:  Fully Operational


Resource Links:


Room 4.022 - Olympus BX51 Upright Microscope Stand with Olympus DP70 CCD camera

Upright widefield microscope with Mercury lightsource and CCD camera. Suited for imaging fixed and live fluorescent samples.

Features Include:

  • Manual X-Y-Z stage
  • Polarised light
  • Differential Interference Contrast (DIC)
  • Optovar (1x, 1.25x, 1.6x, 2x)

Transmitted Light:  Olympus 100W Halogen Lamp

Reflected Light:  Olympus 100W Mercury White Lightsource

Condensor:

Name N.A. W.D. mm Position 1 Position 2 Position 3 Position 4 Position 5 Position 6

Position 7

Position 8

Universal 0.9 - Open Open Open 10x DIC 20x DIC 40x DIC 60x DIC 100x DIC

Objectives:

Position Objective Magnification N.A. Immersion W.D. (mm) Type Type Thread
1 Plan Apochromat 2x 0.08 Dry - -   M27
2 Plan Apochromat 10x 0.40 Dry - -   M27
3 Plan Apochromat 20x 0.75 Dry - -   M27
4 UApo 40x 1.35 Oil - -   M27
5 Plan Apochromat 60x 1.35 Oil - -   M27
6 Plan Apochromat 100x 1.4 Oil - -   M27

Fluorescent Filter Sets:

Position Name Excitation Dichroic Emission Suitable Dyes
1 DAPI - 5060C 377/50 409LP 447/60 DAPI, Hoechst
2 GFP-4050A 466/40 495LP 525/50 EGFP, Alexa Fluor 488
3 mCherry-40LP-A 562/40 593LP 593LP

mCherry, Alexa Fluor 568

4 CFP-2432C 438/24 458LP 483/32 ECFP, mCerulean
5 YFP-2427B 500/24 520LP 542/27 EYFP, mVenus
6 Empty - - - Brightfield

Computer:

Dell PC - 3GHz Xeon Processor - 8GB RAM - 256GB nVidia Quadro GPU

1TB Raid Array

30" LCD Monitor

Software: 

DP Manager, DP Controller

Fluoro 3 - Inverted Olympus IX70

Current Status:  Fully Operational


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Room 2.138 - Olympus IX70 Inverted Microscope Stand with Nikon DS-Fi1 CCD camera

Inverted widefield microscope with Mercury lightsource and CCD camera. Suited for imaging fixed and live fluorescent samples.

Features Include:

  • Manual X-Y-Z stage

Transmitted Light:  Olympus 100W Halogen Lamp

Reflected Light:  XCite 120Q Halogen White Lightsource

Condensor:

Name N.A. W.D. mm Position 1 Position 2 Position 3 Position 4 Position 5 Position 6
Universal LWD 0.45 54 Open PhL Ph1 Ph2 DIC I DIC II

Objectives:

Position Objective Magnification N.A. Immersion W.D. (mm) Type Type Thread
1 Plan Apochromat 4x 0.5 Dry 2 PhL ADL M27
2 Plan Apochromat 10x 0.75 Dry 0.5 Ph1   M27
3 Plan Apochromat 20x 0.75 Dry 0.4 Ph2 ADL M27
4 Plan Apochromat 40x 0.95 Dry 0.2 DIC I   M27
5 Plan Apochromat 63x 1.4 Oil 0.17 DIC II   M27
6 Plan Apochromat 100x 1.4 Oil 0.13 DIC III   M27

Fluorescent Filter Sets:

Position Name Excitation Dichroic Emission Suitable Dyes
1 DAPI - 5060C 377/50 409LP 447/60 DAPI, Hoechst
2 CFP - 2432C 438/24 458LP 483/32 CFP, Cerulean
3 GFP-3000B 488/10 505LP 530/60 GFP, A488
4 mCherry LP 560/20 580LP 585LP mCherry, Cy3
5   - - -  
6   - - -  

Computer:

Dell PC - 3GHz Xeon Processor - 8GB RAM - 256GB nVidia Quadro GPU

1TB Raid Array

30" LCD Monitor

Software: 

NIS Elements

Fluoro 4 - Inverted Nikon Ti-U

Current Status:  Fully Operational


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Room 6.025a - Nikon Ti-U Inverted Microscope Stand with Nikon DS-Fi1 and Qi1 CCD cameras

Inverted widefield microscope with Mercury lightsource and CCD camera. Suited for imaging fixed and live fluorescent samples.

Features Include:

  • Manual X-Y-Z stage

Transmitted Light:  Nikon 100W Halogen Lamp

Reflected Light:  Nikon 100W Mercury White Lightsource

Condensor:

Name N.A. W.D. mm Position 1 Position 2 Position 3 Position 4 Position 5 Position 6
Universal LWD 0.45 54 Open PhL Ph1 Ph2 DIC I DIC II

Objectives:

Position Objective Magnification N.A. Immersion W.D. (mm) Type Type Thread
1 Plan Fluor 4x 0.13 Dry - PhL DL M27
2 Plan Fluor 10x 0.30 Dry - Ph1 DL M27
3 Plan Fluor ELWD 20x 0.45 Dry - Ph1 ADL M27
4 Plan Fluor ELWD 40x 0.60 Dry - Ph2 ADL M27
5 Plan Fluor 100x 1.30 Oil - Ph3 DDL M27
6 - - - - - - - M27

Fluorescent Filter Sets:

Position Name Excitation Dichroic Emission Suitable Dyes
1 DAPI - 5060C 377/50 409LP 447/60 DAPI, Hoechst
2 CFP - 2432C 438/24 458LP 483/32 ECFP, Cerulean
3 GFP-3000B 488/10 505LP 530/60 EGFP, Alexa Fluor 488
4 YFP-2427B 500/24 520LP 542/27 EYFP, mVenus
5 mCherry-40LP-A 562/40 593LP 593LP mCherry Alexa Fluor 568
6 Empty - - - Brightfield

Computer:

Dell PC - 3GHz Xeon Processor - 8GB RAM - 256GB nVidia Quadro GPU

1TB Raid Array

30" LCD Monitor

Software: 

NIS Elements-BR, NIS Elements F

Accessories:

Motorised Z-Focus

Fluoro 5 - Upright Zeiss AxioImager

Current Status:  Fully Operational


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Room 3.011 - Zeiss AxioImage M2.m Microscope Stand with Apotome2 and Axiocam 506 camera

Inverted widefield microscope with Mercury lightsource and CCD camera. Suited for imaging fixed and live fluorescent samples.

Features Include:

Motorised X-Y-Z stage, Apotome2

Transmitted Light:  LED illumination

Reflected Light:  HXP 120c White Lightsource

Condensor:

Name N.A. W.D. mm Position 1 Position 2 Position 3 Position 4 Position 5 Position 6
Motorised Universal 0.9     DIC I DIC II DIC III    

Objectives:

Position Objective Magnification N.A. Immersion W.D. (mm) Type Type Thread
1 EC Plan NeoFluar 5x 0.16 Dry 18.5 DIC   M27
2 Plan Apochromat 10x 0.30 Dry 2 DIC   M27
3 Plan Apochromat 20x 0.80 Dry 0.55 DIC   M27
4 Plan Neofluar 40x 1.30 Oil 0.21 DIC   M27
5 Plan Apochromat 63x 1.40 Oil 0.14 DIC   M27
6 Plan Apochromat 100x 1.40 Oil 0.17 DIC   M27

 

Fluorescent Filter Sets:

Position Name Excitation Dichroic Emission Suitable Dyes
1 DAPI 49 365 395 445/50 DAPI, Hoechst
2 FITC 44 475/40 500 530/50 EGFP, Alexa Fluor 488
3 Cy3 43 545/25 570 605/70 Cy3
4 Texas Red 45 560/40 585 630/75 Tx Red, mCherry, A568
5 Cy5 50 640/30 660 690/50 Cy5, A647
6 Empty - - - Brightfield

 

Computer:

HP Z820 PC - 3GHz Xeon Processor -64GB RAM - 1GB ATI Firepro V5800 GPU

1TB Raid Array

30" LCD Monitor

 

Software: 

Zeiss Zen 2 with Tiling and EDF modules

Accessories: