Imaging biological specimens with the confocal scanning laser microscope/macroscope

S. Damaskinos, A. E. Dixon, K. A. Ellis, W. L. Diehl-Jones

    Research output: Contribution to journalJournal Articlepeer-review

    5 Citations (Scopus)

    Abstract

    A new confocal scanning laser microscope/macroscope (cslm/M) has recently been developed. It combines in one instrument the high resolution capability of a confocal scanning beam microscope for imaging small specimens, with good resolution confocal imaging of macroscopic specimens. Some of its main features include: (a) 0.25 μm lateral resolution in the microscope mode and 5 μm lateral resolution in the macroscope mode; (b) a field of view that can vary from 25 μm × 25 μm to 75,000 μm × 75,000 μm; (c) capability for acquiring large data sets from 512 × 512 pixels to 2048 × 2048 pixels; (d) 0.5 μm depth resolution in the microscope mode and 200 μm depth resolution in the macroscope mode. In this work the cslm/M was used to image whole biological specimens (> 5 m diameter), including insects which are ideal specimens for the macroscope. Specimens require no preparation, unlike scanning electron microscope (SEM) specimens which require a conductive coating. The specimens described in this paper are too large to be imaged in their entirety by a scanning beam laser microscope, however they can be imaged by slower scanning stage microscopes. In the macroscope mode the cslm/M was used to acquire a large number (e.g. 20-40) of confocal image slices which were then used to reconstruct a three-dimensional image of the specimen. High resolution images were collected by the cslm/M by switching to the microscope mode where high numerical aperture (NA) objectives were used to image a small area of interest. Reflected-light and fluorescence images of plant and insect specimens are presented which demonstrate the morphological details obtained in various imaging modes. A process for three-dimensional visualization of the data is described and images are shown.

    Original languageEnglish
    Pages (from-to)493-502
    Number of pages10
    JournalMicron
    Volume26
    Issue number6
    DOIs
    Publication statusPublished - 1995

    Keywords

    • biological specimens
    • confocal
    • fluorescence
    • macroscope
    • microscope
    • three-dimensional visualization

    Fingerprint

    Dive into the research topics of 'Imaging biological specimens with the confocal scanning laser microscope/macroscope'. Together they form a unique fingerprint.

    Cite this