Table of Contents

Location: [Oeiras] Room 0B02
Manufacturer: ZEISS Microscopy
Model: Zeiss Axio Imager 2 with Apotome 2
Nickname: Apotome
Software: Zen 3.1
Year: 2019
SN: 3523001603
 
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Microscope overview

The ZEISS Axio Imager with Apotome 2 is a fully automated upright microscope capable of magnifications from 2.5x to 160x and equipped with two cameras: On the left port a 5MPx CMOS color camera (Axiocam 105 RGB, for brightfield/DIC) and on the top port a high sensitivity 4 Mpx monochrome sCMOS camera (Hamamatsu Flash 4.0v2, for fluorescence). The system has a Colibri 7 excitation light source with 5 LEDs (see table below). The system’s stage, which can accommodate 8 slides simultaneously, is motorized in XYZ, capable of z-stacks, multiple positions or tile and stitching acquisition (thereby allowing acquisition of large fields of view with several hundred megapixels). The Imager is also equipped with ApoTome.2, which allows optical sectioning of fluorescent samples using structured illumination. A grid pattern is placed on the illumination path and is projected and shifted sideways; 3 (or more) images are acquired and between each image the grid is moved 1/3 of its cycle. This movement generates images where only the in-focus light is modulated by the grid pattern while out-of-focus light is largely unaffected by the grid. By subtracting pairs of these images, the system removes out-of-focus light and the grid pattern, thus creating an optical section of the sample.

System components

LEDs

Line Wavelength Power (with 10x)
UV 385 nm ~ 160* mW
B 475 nm ~ 120 mW
G 555 nm ~ 20 mW
Y 590 nm ~ 14 mW
R 630 nm ~ 44 mW

* don't exceed 5% power as this line bleaches samples quickly.

Filtersets

Filterset Excitation Dichroic Emission Reference
Blue 370-410 nm 420 nm 430-470 nm FS96HE
Green 450-490 nm 493 nm 500-550 nm FS38HE
Orange-Red 540-552 nm 580 nm > 590 nm FS15
Red 625-655 nm 660 nm 665-715 nm FS50
DIC - - - -
Orange 575-600 nm 605 nm 612-682 nm FS64HE
QUAD 370-400 nm 405 nm 410-440 nm FS90HE

Objectives

Magnification Model Immersion NA WD (mm) Reference
2.5x Plan-Neofluar air 0.085 8.8 420320-9902-000
10x Plan-Neofluar air 0.3 5.5 420340-9900-000
20x Plan-Apochromat air 0.8 0.55 420650-9902-000
40x Plan-Apochromat air 0.95 0.25 420660-9970-000
40x Plan-Neofluar oil 1.3 0.21 420462-9900-000
100x Plan-Apochromat oil 1.4 0.17 420792-9900-000

Camera

Model Frame Size Pixel Size (µm) Quantum Efficiency
Hamamatsu ORCA-Flash4.0 V2 2048 x 2048 13.3 x 13.3 70 %
ZEISS Axiocam 105 color 2048 x 2048 2.2 x 2.2 -

Microscope Turn On Procedure

Microscope Turn Off Procedure

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