Life Science Applications of Light Microscopes

Axio Imager from Carl Zeiss is a more flexible imaging platform that is suitable for a myriad of applications.

The modular architecture of Axio Imager 2 enables the users to employ a more suitable technology that supports their application, and grows with their performance requirements. This article discusses the various life science applications of Axio Imager.

Pathology

Axio Imager A2 with LED illumination, the coded stand with Fixed Kohler illumination is suitable for pathology. Together with EC Plan-NEOFLUAR or Plan-APOCHROMAT objectives, the platform is a standard tool for histological evaluation.

The LED illumination is durable and economical as it consumes small amount of energy and does not require maintenance or adjustment.

It delivers high-quality images, for example with the typical H.-E-, DAB- or Azan staining methods. It provides homogenous light quality and incredible image presentation over the entire intensity range thanks to its constant color temperature (see Figure 1).

Salivary gland: azan staining; Orange: cytoplasm, Red: nuclei, Blue: collagen. Objective: Plan-APOCHROMAT 20x/0.8

Figure 1. Salivary gland: azan staining; Orange: cytoplasm, Red: nuclei, Blue: collagen. Objective: Plan-APOCHROMAT 20x/0.8

Human Genetics

Genome analysis is a standard tool used in human genetics to diagnose diseases caused by a mutation in genetic material through the acquisition and analysis of karyograms in transmitted light brightfield.

The Fluorescence In Situ Hybridization (FISH) technique determines the gene loci on the chromosomes depending on the DNA probes utilized to identify any deviation from the healthy condition. The Axio Imager delivers complete support for this application. The object field is illuminated homogenously for all colors by the apochromatically corrected IC2S beam path. The stray light in the illumination and imaging beam path is eliminated by the integrated light traps (Figure 2).

Multi-color FISH preparation. Objective: Plan-APOCHROMAT 63x/1.4 Oil

Figure 2. Multi-color FISH preparation. Objective: Plan-APOCHROMAT 63x/1.4 Oil

The 10-position reflector turret for Axio Imager.Z2 andAxio Imager.D2 as well as the 6-position reflector turret for Axio imager A2 and Axio Imager.M2 enable rapid multi-channel image acquisition, the cornerstone for FISH analyses. Control with AxioVision from Carl Zeiss or MetaCyte from MetaSystems simplifies such such complex applications.

Histology

Histology and anatomy require optimal image resolution, rapid and accurate relocalization of diagnostically conclusive spots in the specimen, and perfect color presentation in the documentation of details and overviews. The combination of the EC Plan-NEOFLUAR and Plan-APOCHROMAT objectives and motorized stages is ideally customized for this application (Figure 3).

Histological section – brightfield. Red: Anti-CD. Blue: nuclear counterstaining. Objective: Plan-APOCHROMAT 63x/1.4 Oil

Figure 3. Histological section – brightfield. Red: Anti-CD. Blue: nuclear counterstaining. Objective: Plan-APOCHROMAT 63x/1.4 Oil

Cell Biology

The analysis of sub-cellular compartments, including the cell nucleus, vesicles, and mitochondria, or dynamic processes, including mobility, motility, and cell division, demands special requirements on the corresponding microscope systems. Axio Imager enables brilliant DIC, darkfield applications, phase contrast, and optical sections with ApoTome in addition to fluorescence at the highest resolution (Figure 4). It is easy to combine DIC and fluorescence with the motorized Axio Imager.Z2 stand.

Brain section (rat) – multichannel image with ApoTome. Green: GFP-labeled astrocytes. Blue: cell nuclei (DAPI). Objective: Plan-APOCHROMAT 20x/0.6. E. Fuchs, S. Bauch, DPC, Göttingen, Germany

Figure 4. Brain section (rat) – multichannel image with ApoTome. Green: GFP-labeled astrocytes. Blue: cell nuclei (DAPI). Objective: Plan-APOCHROMAT 20x/0.6. E. Fuchs, S. Bauch, DPC, Göttingen, Germany

Neurobiology

In neurobiology, different samples ranging from individual cells and thin to thicker brain sections to entire brains are used to obtain meaningful results. Axio Imager is the suitable instrument for this application as it provides superior image quality in brightfield, high-contrast DIC images for very thin sections, and fluorescence, high-resolution DIC for thick preparations. The motorization of all key components and the utilization of the motorized DIC turret on Axio Imager.Z2 enable the storage of all key settings for reproducible imaging and succeeding image analysis.

Developmental Biology

The highest performance of color fidelity, contrast, and resolution is crucial irrespective of the animal model utilized. Axio Imager delivers the superior optical resolution, ideal homogeneous illumination in the general transmitted light contrast methods, and unprecedented sample protecting fluorescence illumination with optimal signal-noise ratio to provide brilliant image quality.

It enables the imaging of the processes to be explored at high spatial and temporal resolution and their analysis with different AxioVision Modules. The motorization of Axio Imager M- and Z-stands facilitates efficient and reproducible imaging (Figures 5-7).

Interphase

Figure 5. Interphase

Metaphase

Figure 6. Metaphase

Telophase

Figure 7. Telophase

HeLa cells – mitosis stages. Red: Alexa Fluor 594-DM1-alpha. Green: Alexa Fluor 488-Mad2. Blue: DNA (DAPI). Objective: EC Plan-NEOFLUAR 100x/1.3 Oil. H.Y. Li, Y. Xheng, HHMI & CIW, Dept. Embryology, MD, USA

This information has been sourced, reviewed and adapted from materials provided by Carl Zeiss Microscopy GmbH.

For more information on this source, please visit Carl Zeiss Microscopy GmbH.

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