Zeiss Axio Observer 7 with deconvolution

 

The AMI widefield microscope is a microscope with a wide array of useful functions. Combined with an incubator to control both temperature and carbon dioxide, the AMI widefield can image live samples for extended periods of time. Any microscopic sample can be viewed with this microscope, ranging from cultured mammalian cells, bacterial cells, and single cell ciliates to Drosophila embryos and migrating immune cells. Experiments that take advantage of the low photo-exposure imposed on the sample—as well as the relatively high speed of widefield microscopes—can yield significant information on the processes of life at the micron scale.

 

The benefits and versatility of the AMI widefield can be further improved by the deconvolution of captured images. Where suitable, this computational method improves the clarity of the images captured, allowing researchers to gain slightly more information on structures and biological processes that may not have been visible prior to deconvolution. 

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Instrument specifications

• Inverted microscope configuration

• Auto focus for time-lapse acquisitions of live samples in aqueous media

• Contrast methods:

  • bright-field transmitted light

  • phase contrast

  • differential interference contrast (DIC)

  • fluorescence

• Temperature-, relative humidity, and CO₂-controlled incubation

• Objectives:​

  • 100× 1.4 NA oil-immersion with phase contrast

  • 63× 1.4 NA oil-immersion with DIC

  • 40× 1.2 NA water-immersion with correction collar

  • 40× 0.95 NA with correction collar

  • 20× 0.35 NA with phase contrast

  • 10× 0.25 NA with phase contrast

• Light source:

  • Halogen lamp for transmitted light

  • LED for fluorescence

    • ​735nm ■

    • 630nm ■

    • 590nm ■

    • 555nm ■

    • 475nm ■

    • 430nm ■

    • 385nm ■

• Emission filters:​

  • Simultaneous quad-band pass

    • ​410-440nm ■

    • 499-529nm ■

    • 567-617nm ■

    • 659-759nm ■

  • Simultaneous tri-band pass

    • 455-489nm ■

    • 543-568nm ■

    • 608-753nm ■

  • ​Simultaneous quad-band pass

    • 413-438nm ■

    • 501-552nm ■

    • 619-657nm ■

    • 766-804nm ■

• Photometrix Prime 95B sCMOS monochrome camera

• Automated stage control with piezo X-, Y-, and Z-control

 

Instrument capabilities

• Resolution limit: ~200nm xy; 1100nm z

• Speed limit: down to 30ms for single frame and 2.5s for a 10μm section

• Depth: down to 25μm

• Field of view: dependent on objective

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Instrument uses​

• Live-cell imaging

• Automated stage movement

• Tiling and panorama (for large fields of view)

• Extended depth-of-field (transmitted light)

• Cell migration and movement

• Organelle tracking

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