Widefield Microscopy

This automated upright widefield microscope has a motorized stage and can be used to visualize fixed preparations that have been labeled using any of a wide variety of fluorophores and dyes. It has two cameras: a monochrome camera for imaging fluorescence and a color camera for imaging brightfield and DIC (Zeiss Axiocam 503: 1936 X 1460 pixels, 4.54 µm X 4.54 µm per pixel, monochrome sensitivity ~400nm – 1000nm; color sensitivity ~400nm – 720nm, RGB Bayer color filter mask).  It is also equipped with Zeiss’s ApoTome.2, which uses structured illumination to generate optical sections with any of the objectives. The motorized stage makes the AxioImager.Z2 ideal for conducting systematic scans or for generating tiled images. Hardware is controlled by Zeiss Zen Blue software, which also incorporates some image processing options.

Zeiss AxioImagerZ2M with motorized stage

Fluoresence Filters

Fluorophore

Excitation

Beam split

Emission

DAPI

AT350/50x

T400LP

ET460/50m

GFP/FITC

ET470/40x

T495LP

ET525/50m

TRITC

ET545/40x

T570LP

ET620/60m

Cy5

ET620/60x

T660LPXR

ET700/75m

Cy7

ET710/75x

T760LPXR

ET810/90m

CFP*

ET436/20x

T455LP

ET480/40m

YFP*

ET500/20x

T515LP

ET535/30m

Spectrum Orange*

HQ548/20x

Q565LP

HQ580/14m

* Available on request

Objectives

Magnification

Immersion

N.A.

10X

air

0.3

20X

air

0.8

40X

air

0.75

40X

oil

1.3

63X

oil

1.4

100X

oil

1.4

Brightfield Protocol
Fluorescence Protocol 

This automated upright widefield microscope is equipped to capture images of fixed preparations that have been labeled with a wide variety of fluorophores and dyes. It is equipped with both a monochrome camera for imaging fluorescence (Zeiss Axiocam 503 mono; 1936 X 1460 pixels, 4.54 µm X 4.54 µm per pixel, sensitivity ~400nm –1000nm) and a color camera for imaging brightfield and DIC (Zeiss Axiocam MRc; 1300 X 1030 pixels, 6.7 µm X 6.7 µm per pixel, sensitivity 400nm – 710, Bayer color filter mask). It is also equipped with Zeiss’s ApoTome.2, which uses structured illumination to generate optical sections with all but the two lowest magnification objectives (2.5X and 5X). Hardware is controlled by Zeiss Zen Blue software, which also incorporates some image processing options.

Zeiss AxioImagerZ2

Fluorescence Filters

Fluorophore

Excitation

Beam split

Emission

DAPI

AT350/50x

T400LP

ET460/50m

GFP

ET470/40x

T495LPXR

ET525/50m

Texas Red

ET560/40x

T585LPXR

ET630/75m

Cy5

ET620/60x

T660LPXR

ET700/75m

CFP

ET436/20x

T455LP

ET480/40m

YFP

ET500/20x

T515LP

ET535/30m

Spectrum Orange*

HQ548/20x

Q565LP

HQ580/14m

* Available on request

Objectives

Magnification

Immersion

N.A.

2.5X*

air

0.075

5X*

air

0.15

10X

air

0.75

20X

air

0.75

40X

oil

1.3

63X

oil

1.4

100X

oil

1.4

Brightfield Protocol
Fluorescence Protocol

This automated upright microscope controlled by Leica Application Suite: Advanced Fluorescence software has a Leica DFC350 FX camera for capturing fluorescent dyes in multicolor fluorescence applications.  It has air lenses up to the 63x range, so it provides a simple, clean, and fast solution for imaging standard slides with fixed cells. It can perform transmitted light imaging techniques such as polarized light and DIC, and it utilizes a motorized stage that comes in handy for automated montage imaging. 

Leica DM6000

Fluorescence Filters

Fluorophore

Excitation

Beam split

Emission

DAPI

BP 360/40

400

BP 470/40

GFP

BP 470/40

500

BP 525/50

FITC

BP 480/40

505

BP 527/30

Rhodamine

BP 546/12

565

BP 600/40

Cy3

BP 545/30

565

BP 610/75

Cy5

BP 620/60

660

BP 700/75

Objectives

Magnification

Immersion

NA

1.25x

Air

0.04

5x

Air

0.15

10x

Air

0.3

20x

Air

0.5

40x

Air

0.85

63x

Air

0.9

100x

Oil

1.4

User Protocol

This automated inverted microscope has multiple features to enable long-term time lapse imaging experiments: a LiveCell enclosed chamber that permits temperature, C02, and humidity control; adapters for well plates, dishes and slides; and a Ludl motorized stage and focus motor that can be used to mark multiple sites in xyz. Images are captured with a Q Imaging Retiga 2000R cooled CCD camera; 1600 X 1200 pixels, 7.4µm X 7.4µm per pixel, sensitivity is high and linear through ~550nm and then decreases linearly between 600nm to ~900nm. Exposure to light is controlled by Uniblitz shutters. The system is controlled by MetaMorph software. An on-site tissue culture incubator is available to house cells prior to imaging. Typical uses for this system include cell tracking, neurite growth, cell motility, wound healing, and Fura-2 imaging.

At left, Olympus Live Cell System. At right, images of the same specimen at increasing magnification taken by Crystal Pristell

Fluorescence Filters

Fluorophore

Excitation

Beam split

Emission

DAPI

D350/50x

83100bs

S450/36m

FITC

S492/18x

83100bs

S530/35m

Texas Red

S572/23x

83100bs

S630/60m

CFP

S430/25x

51017bs

S470/30m

YFP

S510/20x

51017bs

S535/30m

Cy5

S655/20x

86020bs

S710/75m

FURA2*

ET340x ET380x

T400LP

ET510/80m

Objectives

Magnification

Immersion

N.A.

5X

air

0.15

10X

air

0.3

20X

air

0.45

40X

air

0.6

60X

oil

1.42

20X*

air

0.5

User Protocol

This automated upright microscope controlled by Zeiss AxioVision software has a Zeiss AxioCam MRc camera for capturing color bright field images. It is also equipped for DIC, plane polarized and circularly polarized light microscopy.

Axioplan2 microscope is equipped with optics for brightfield (left), plane polarized (center) and circularly polarized (right) light microscopy. Lamb hoof stained with picosirius red by Virginia Gillespie.

Fluorescence Filters

Fluorophore

Excitation

Beam split

Emission

DAPI

AT350/50x

T400LP

ET460/50m

GFP

ET470/40x

T495LPXR

ET525/50m

Texas Red

ET560/40x

T585LPXR

ET630/75m

Cy5

ET620/60x

T660LPXR

ET700/75m

CFP

ET436/20x

T455LP

ET480/40m

YFP

ET500/20x

T515LP

ET535/30m

Spectrum Orange*

HQ548/20x

Q565LP

HQ580/14m

Objectives

Magnification

Immersion

NA

10x

air

0.3

20x

air

0.75

40x

air

0.75

40x

oil

1.3

63x

oil

1.4

100x

oil

1.4

*available on request

Brightfield Protocol
Fluorescence Protocol
Polarized Light Protocol

This system is an Olympus (stereo) microscope system, equipped with a DP80 dual chip color and monochrome camera (1360 x 1024), making it capable of both fluorescence and full color brightfield imaging, with accurate pixel-to-pixel correspondence. The system is controlled by cellSens software. The stereoscope is also capable of manual montage acquisition and stitching for high resolution images of large areas.  In addition, the MVX10 body includes a 0.63x to 6.3x adjustable zoom factor, to easily tailor magnification to individual specimens.

At left, Olympus Macroscope. At right, skeletal prep from an E16.5 mouse stained with Alizarin Red (bone) an Alcian Blue (cartilage) by Jianyun Yan.

Fluorescence Filters

Fluorophore

Excitation

Beam split

Emission

DAPI

AT350/50x

T400LP

ET460/50m

GFP

ET470/40x

T495LPXR

ET525/50m

DsRED

ET545/25x

T565LPXR

ET605/70m

CY5*

ET620/60x

T660LPXR

ET700/75m

CFP*

ET438/20X

T450LPXR

ET518/20m

Objectives

Magnification

Immersion

NA

0.63x*

air

0.15

1x

air

0.25

2x

air

0.50

*available on request

User Protocol