AxioImager DIC/Fluorescence Scope | Neuroscience Microscopy Service

The AxioImager is a motorized, upright microscope equipped for both transmitted light (DIC) and fluorescence microscopy.

This microscope is perfect for Prof. Stephen Smith's Array Tomography technique, as well as general-purpose widefield microscopy.

Fast Facts

Fully automated microscope allowing tiled images, z-stacks and time-lapse imaging
Dual CCD cameras: monochrome and color
Four-color multi-channel fluorescence with monochrome CCD
Histological stains with color CCD camera.

Technical Specifications

Z drive	DC motorized Z-drive with optoelectronic coding, resolution 10 nm, reproducibility ±10 nm
XY stage	Piezo-driven XY scanning stage with linear encoders, travel range: 225 × 85 mm, maximum speed: 100 mm/s, resolution: 0.2 μm, reproducibility: < 1 μm, absolute accuracy: ±5 μm
Transmitted light	12 V/100 W halogen light source, manual condenser, 0.9 NA, brightfield/DIC/phase
Optovar	Manual 3-position tube lens: mag. 1.0×, mag. 1.6×, Bertrand lens
Fluorescence equipment	EXFO X-Cite 120 illumination source, coupled to scope with liquid light guide, motorized 6-position reflector turret with DIC cube and five fluorescence filter cubes (see below).
Camera	Zeiss AxioCam HRm Rev. 3. Full specifications below.
Software	Zen 2.3 with z-stack, automated tiling, autofocus and multi-positional imaging

Objectives (each with DIC slider)
mag.	type	NA	working distance (mm)	pdf file
2.5×	Plan Neofluar	0.075	9.5	click here
10×	Plan Apochromat	0.45	2.0	click here
20×	Plan Apochromat	0.8	0.55	click here
40×	Plan Apochromat	0.95	0.25	click here
63× (oil)	Plan Apochromat	1.4	0.19	click here

Fluorescence filter sets
description	excitation peak/FWHM (nm)	dichroic 50% trans. (nm)	emission peak/FWHM (nm)	pdf file
DAPI	365/50	395	445/50	DAPI.pdf
eGFP	470/40	495	525/50	eGFP.pdf
Cy3	550/25	570	605/70	Cy3.pdf
mCherry	560/40	585	630/75	mCherry.pdf
Cy5	640/30	660	690/50	Cy5.pdf

Camera Specifications: AxioCam HRm Rev. 3

Sensor

Sony ICX 285, progressive readout

CCD Basic resolution

1388 × 1040 = 1.4 megapixels

Pixel size

6.45 μm (h) × 6.45 μm (v)

Sensor size

Chip area 8.9 mm × 6.7 mm (equivalent 2/3 inch)

Spectral range

Approximately 350 nm–1000 nm with BK-7 protection glass without IR filter (BG40 can be inserted if necessary).

NIR mode

Mode for higher sensitivity in near IR

Dynamic range

Typical > 1:2200 at 25 MHz with < 7.7 e readout noise
Typical > 1:2500 at 12.5 MHz with < 6.8 e readout noise

Full well

Typical 17,000 e

Dark current

Typical 0.7 e/pixel/s, optional dark current compensation for maximum low light performance at long exposure times

Resolution improvement

Microscanning technology enables a configurable image resolution beyond the basic sensor resolution

Selectable resolution

       H × V (single shot)
   276 × 208  (binning 5 × 5)
   346 × 260  (binning 4 × 4)
   462 × 346  (binning 3 × 3)
   694 × 520  (binning 2 × 2)
1388 × 1040  (no binning)

       H × V (microscanning)
2776 × 2080  (microscanning mode)
4164 × 3120  (microscanning mode)

Live image frame rates

H × V	Mode / Binning	Max. frame rate (25 MHz)
1388 × 1040	Slow / 1	12 images/s
694 × 520	Middle / 2	21 images/s
462 × 346	Fast / 2	31 images/s

Readout of sub-frames (ROI)

Random definition of regions of interest (ROI) on the sensor enables further increase of achievable frame rates

Signal amplification

Analog: 2×, digital: 32×

Digitization

Two switchable readout speeds offer different depths of digitization:
High Quality: 14 bit / 12.5 MHz
High Speed: 12 bit / 25 MHz

CCD cooling

Single stage Peltier cooling, regulated

Integration time

1 ms up to several minutes

For an exploded diagram overview of this microscope, axioimagersystemoverview.pdf
For the complete AxioImager product brochure, zeissaxioimagerbrochure.pdf