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Instrument Types

Different microscopes have different uses.

 

Wide Field

Wide field microscopes illuminate the whole field of view at one time.  Imaging is fast, but out of plane information is included as blur.  There are computational methods that can reduce, but not eliminate the blur (e.g., deconvolution and extended depth of focus).

 

The Cell Imaging Core has wide field imaging systems.  These include:

  • EVOS Auto Color and Fluor
  • Nikon Automated Widefield
  • DeltaVision Widefield
  • Axio Scan.Z1 Slide scanner (details)
  • Axioscan 7 Slide scanner

Scanning Confocal

Scanning confocal microscopes illuminate a spot at one time and scan across the field of view.  In a confocal system, there is also a pinhole to limit out of focal plane blur.  These are slower than wide field, but provide better resolution, especially in Z.  Further, they can be used to image the sample in 3 dimensions.

Spinning Disk Confocal

Spinning disk confocal microscopes use several spots at one time to reduce the imaging time, making them suitable for live cell imaging.

Multi-Photon

2-photon (and multi-photon) microscopes rely on 2 or more simultaneous lower energy photons to stimulate fluorescence.  One of the main advantages of this method is that the lasers can penetrate deeper into the sample.  It does require a more powerful laser and working in the IR.  Applications include samples up to a few mm thick and intravital microscopy.

STED

STimulated Emission Depletion uses a laser to deplete emission around the focal spot.  This provides much higher lateral resolution.  This is one of a few super-resolution microscopy methods.

Airy Scan

In an Airy scan microscope, multiple detectors are used to interrogate the spatial blur or Airy pattern.  This additional information is then used to reduce the blur, achieving higher resolution than normal confocal.

Structured Light

If several images are made with known patterns of light, the images can be combined to create a higher resolution image.

TIRF

Total Internal Reflection Fluorescence microscopy achieves thin Z sectioning at the expense of only being able to image the first thin (<200nm) layer of the sample.  It does have a high signal to noise ratio because there is very little out of plane fluorescence.  This makes the method useful for for events in cellular surfaces.

 

The Cell Imaging Core has several confocal imaging systems:

  • Zeiss 700 Confocal Microscope (details)

 

 

 

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