|Widefield||Spinning disk||TIRF||Lattice LightSheet|
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Widefield microscopy illuminates the whole sample with light and collects the image onto a camera. Widefield microscopy provides good images of thin samples. For thicker samples, the lack of optical sectioning means a widefield microscope might not be the best option but images can be improved with deconvolution post-processing to increase the resolution and reduce noise.
Spinning disk confocal
Spinning disk confocal uses multiple points of laser light to illuminate the sample and multiple pinholes to eliminate out-of-focus light. Spinning disks are used for high-speed, high resolution, optically sectioned imaging. They do, however, suffer from reduced background rejection compared to point-scanning confocals in thick samples (if you are interested in using a point-scanning confocal, get in contact with the SLS Imaging Suite).
Total internal reflection fluorescence microscopy (TIRFM) uses an evanescent wave to illuminate the bottom 100 nm of the sample. It gives the best available axial resolution but is only suitable for 2D imaging proximal to the coverslip.
Lattice light sheet microscopy
Light sheet microscopy is a completely different way of imaging, the excitation and detection objective are orthogonal to each other. This gives increased optical sectioning with minimal photobleaching. Advanced beam-shaping allows the lattice light sheet to go at high speed with high resolution while reducing phototoxicity. If you are interested in using the Lattice LightSheet please contact Helena Coker.