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ZEISS Apotome 3 with Axio Observer
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ZEISS Apotome 3 Optical Sectioning in Widefield Fluorescence Microscopy

Optical sectioning with ZEISS Apotome 3 allows you to efficiently minimize out-of-focus light. Create crisp images and 3D renderings, even of thicker specimen, while your microscope remains just as easy to operate as always. Go one step further with Apotome Plus and achieve confocal-like image quality with your widefield microscope.

  • Reliable optical sections thanks to structured illumination
  • Based on linear approaches and peer-reviewed algorithms
  • 3D structural analysis with resolution down to 180 nm

ZEISS Apotome 3 with Apotome Plus

See How It Works

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ZEISS Apotome 3 with Apotome Plus

See How It Works

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Cos7 cells (nuclei stained with Hoechst, tubulin with Alexa 488 and Phalloidin with Alexa 568) imaged with Plan Apochromat 63x/1.4.

Reliable Optical Sections

Under Varying Experimental Conditions

Apotome 3 significantly increases the axial resolution compared to conventional fluorescence widefield microscopy: You obtain optical sections that allow 3D rendering, even from thick specimens. Three grids of different geometries give you the best resolution for each objective. You can focus on your experiment as the ideal illumination structure is automatically selected, always resulting in high-contrast optical sections.

 

Caption: Cos7 cells (nuclei stained with Hoechst, tubulin with Alexa 488 and Phalloidin with Alexa 568) imaged with Plan Apochromat 63x/1.4.

Cortical neurons (left: Widefield; right: Apotome 3). Courtesy of L. Behrendt, Leibniz-Institute on Aging – Fritz-Lipmann-Institut e.V. (FLI), Germany.
Cortical neurons (left: Widefield; right: Apotome 3). Courtesy of L. Behrendt, Leibniz-Institute on Aging – Fritz-Lipmann-Institut e.V. (FLI), Germany.

Peer-reviewed Algorithms

Linear Approaches for True Optical Sections

Purely software-based methods require either prior knowledge of the sample (AI based methods) or rely on complex algorithms that have not been peer-reviewed. Users must trust that these black-box solutions do not falsify information when “enhancing” the image. ZEISS Apotome 3 uses the information from the structured illumination combined with documented algorithms to create a crisp optical section you can trust.

Caption: Cortical neurons (left: Widefield; right: Apotome 3). Courtesy of L. Behrendt, Leibniz-Institute on Aging – Fritz-Lipmann-Institut e.V. (FLI), Germany.

35 μm sagittal section of adult mouse brain, imaged with ZEISS Axio Observer and ZEISS Apotome, processed with Apotome Plus. Sample courtesy of University of California, Davis / NIH NeuroMab Facility.

Sagittal Section of Adult Mouse Brain

Sample courtesy of University of California, Davis / NIH NeuroMab Facility.
Sample courtesy of University of California, Davis / NIH NeuroMab Facility.

Confocal-like Image Quality

180 nm Resolution with Apotome Plus

Resolve details that were not visible before with your widefield microscope: With Apotome Plus, you can obtain structural information at lateral resolution down to 180 nm. The combination of structured illumination with state-of-the-art image processing substantially improves the signal-to-noise ratio and resolution in x, y, and z.

Caption: 35 μm sagittal section of adult mouse brain, imaged with ZEISS Axio Observer and ZEISS Apotome, processed with Apotome Plus. Sample courtesy of University of California, Davis / NIH NeuroMab Facility.

Free Choice of Light Source and Dyes

Free Choice of Light Source and Dyes

It’s Your Decision, Not the Technology’s

Your experiments often evolve in complexity and requirements. That’s why you need adaptable equipment. Use Apotome 3 with metal halide lamps, economic white light LEDs, or a gentle, multi-color LED light source of the ZEISS Viluma illumination system. Whether you work with DAPI, Alexa488, Rhodamin, Cy5, or with vital dyes such as GFP or mCherry – Apotome 3 adapts to your fluorophores and light source, creating the sharp and brilliant images you expect.

Your Flexible Choice of Components

Customize Your Microscope by Combining Apotome 3 with the Accessories Required for Your Research
  • Microscope​

    Microscope​

    • Axio Observer series (inverted research microscope)​​
    • Axio Imager 2 series (upright research microscope)​​
    • Axio Zoom.V16 (zoom microscope)​​
    • Simple upgrading of existing systems
  • Recommended objective classes​

    Recommended objective classes​

    • C-Apochromat​​
    • Plan-Apochromat​​
    • EC Plan-Neofluar
  • Illumination: Viluma Family

    Illumination

    • Viluma 5/7/9 (LED)​
    • Xylis LED (white light LED)​​
    • HBO (mercury vapor lamp)​​
    • HXP 120 C (metal halide)
  • Cameras: Axiocam 820 mono

    Cameras

    • Monochrome, low-noise ZEISS Axiocam​ camera models​​
    • Selected 3rd-party cameras
A: Widefield image. B – D: Raw images acquired with different grid positions. E: Resulting image; out-of-focus light is efficiently removed by the structured illumination.
A: Widefield image. B – D: Raw images acquired with different grid positions. E: Resulting image; out-of-focus light is efficiently removed by the structured illumination.

A: Widefield image. B – D: Raw images acquired with different grid positions. E: Resulting image; out-of-focus light is efficiently removed by the structured illumination.

A: Widefield image. B – D: Raw images acquired with different grid positions. E: Resulting image; out-of-focus light is efficiently removed by the structured illumination.

The Apotome 3 Operation Principle

Apotome 3 uses a grid to generate a pattern of intensity differences. If out-of-focus light is present at a certain region of the sample, the grid becomes invisible. After the fluorescence of a grid position is acquired, the grid moves to the next position. A true optical section with higher contrast and resolution is calculated.

ZEISS Apotome 3 at Work

  • Cortical neurons stained for DNA, microtubules and microtubule-associated proteins.

    Cortical neurons stained for DNA, microtubules and microtubule-associated proteins. Courtesy of L. Behrendt, Leibniz-Institute on Aging – Fritz-Lipmann-Institut e.V. (FLI), Germany.
  • Transgenic zebrafish larvae at 4 days post fertilization staining for: Glial fibrillary acidic protein, acetylated Tubulin, GFP and DNA. Embedded in 1.2% low melt agarose.

    Transgenic zebrafish larvae at 4 days post fertilization staining for: Glial fibrillary acidic protein, acetylated Tubulin, GFP and DNA. Embedded in 1.2% low melt agarose. Courtesy of H. Reuter, Leibniz-Institute on Aging – Fritz-Lipmann-Institut e.V. (FLI), Germany.
  • Sagittal section of adult mouse brain, imaged with ZEISS Axio Observer and ZEISS Apotome, processed with Apotome Plus.

    Sagittal section of adult mouse brain, imaged with ZEISS Axio Observer and ZEISS Apotome, processed with Apotome Plus. Sample courtesy of University of California, Davis / NIH NeuroMab Facility.
  • Bovine pulmonal arterial endothelial (BPAE) cells having nuclei stained with DAPI, F-actin with Alexa 488 phalloidin and mitochondria with MitoTracker Red CMXRos. Compared to the widefield image, Apotome removes out of focus light creating a crisp optical section. Apotome Plus further improves image quality and resolution resolving even finer structures.

    Bovine pulmonal arterial endothelial (BPAE) cells having nuclei stained with DAPI, F-actin with Alexa 488 phalloidin and mitochondria with MitoTracker Red CMXRos. Compared to the widefield image, Apotome removes out of focus light creating a crisp optical section. Apotome Plus further improves image quality and resolution resolving even finer structures.

ZEISS Apotome 3 at Work

  • Cortical neurons stained for DNA, microtubules and microtubule-associated proteins.

    Cortical neurons stained for DNA, microtubules and microtubule-associated proteins. Courtesy of L. Behrendt, Leibniz-Institute on Aging – Fritz-Lipmann-Institut e.V. (FLI), Germany.
  • Transgenic zebrafish larvae at 4 days post fertilization staining for: Glial fibrillary acidic protein, acetylated Tubulin, GFP and DNA. Embedded in 1.2% low melt agarose.

    Transgenic zebrafish larvae at 4 days post fertilization staining for: Glial fibrillary acidic protein, acetylated Tubulin, GFP and DNA. Embedded in 1.2% low melt agarose. Courtesy of H. Reuter, Leibniz-Institute on Aging – Fritz-Lipmann-Institut e.V. (FLI), Germany.
  • Sagittal section of adult mouse brain, imaged with ZEISS Axio Observer and ZEISS Apotome, processed with Apotome Plus.

    Sagittal section of adult mouse brain, imaged with ZEISS Axio Observer and ZEISS Apotome, processed with Apotome Plus. Sample courtesy of University of California, Davis / NIH NeuroMab Facility.
  • Bovine pulmonal arterial endothelial (BPAE) cells having nuclei stained with DAPI, F-actin with Alexa 488 phalloidin and mitochondria with MitoTracker Red CMXRos. Compared to the widefield image, Apotome removes out of focus light creating a crisp optical section. Apotome Plus further improves image quality and resolution resolving even finer structures.

    Bovine pulmonal arterial endothelial (BPAE) cells having nuclei stained with DAPI, F-actin with Alexa 488 phalloidin and mitochondria with MitoTracker Red CMXRos. Compared to the widefield image, Apotome removes out of focus light creating a crisp optical section. Apotome Plus further improves image quality and resolution resolving even finer structures.

Related Applications

Cell Imaging with Fluorescence Microscopy

With different technologies available that balance sensitivity, resolution and fluorophore requirements, explore your options from widefield to deconvolution, confocal to super-resolution.

Looking Deep Into Chemically Cleared Brain & Thick Tissues

With multiple systems optimized for cleared tissues, find the right technology that balances sample size and the level of detail you wish to resolve.

Downloads

  • Preview image of ZEISS Apotome 3

    ZEISS Apotome 3

    Optical sectioning in fluorescence imaging for your widefield microscope
    7 MB
    Download
  • Preview image of ZEISS Apotome 3 - Flyer

    ZEISS Apotome 3 - Flyer

    Hardware-based, Quantitative Optical Sectioning with Well Documented Algorithms
    1 MB
    Download

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