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Confocal Microscopy

Confocal microscopy using DLP for high resolution images 3-D reconstructions, capable of isolating and collecting a plane of focus from within a sample.

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Design Considerations

Confocal Microscopy DiagramConfocal microscopy is a valuable tool for obtaining high resolution images and 3-D reconstructions. The most important feature of a confocal microscope is the capablity of isolating and collecting a plane of focus from within a sample, thus eliminating the out of focus "haze" normally seen with a fluorescent sample. Fine detail is often obscured by the haze and cannot be detected in a non-confocal, fluorescent microscope.

Using DLP® technology in this application allows users to easily change observation conditions and removes unwanted vibration that effects viewing. DLP® technology serves two purposes; scanning and configuring the illumination and detection pinhole array. The illumination pinhole is created by turning "on" one micro mirror while surrounding mirrors remain in the "off" state. Thus only light reflected from this one micromirror will pass through the optical system. The mirror’s image in the objective lens serves as the illumination pinhole focused on the object. Light reflected "off" the sample at this pin point is then focused back onto the same micromirror of the DMD.

Normally when an object is imaged in the fluorescence microscope, the signal produced is from the full thickness of the specimen which does not allow most of it to be in focus to the observer. The confocal microscope eliminates this out-of-focus information by means of a confocal "pinhole" situated in front of the image plane which acts as a spatial filter and allows only the in-focus portion of the light to be imaged.

To image the entire field of view, a mosaic is configured by shifting the on and off mirrors in a time varying pattern that covers the entire field of view. Using a horizontal scan and shifting the pattern by one mirror several times for each vertical location, the entire sample field is scanned. A CCD camera is then used to acquire one integrated continuous image as the DMD performs scanning.

Application Notes (1)

Title Abstract Type Size (KB) Date Views
HTM 8 KB 19 Aug 2008 3617
    

Selection and Solution Guides

Selection Guides (1)

Title Abstract Type Size (KB) Date Views
PDF 9.03 MB 02 Apr 2013 13116

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