Orthogonal‐plane fluorescence optical sectioning: Three‐dimensional imaging of macroscopic biological specimens

  title={Orthogonal‐plane fluorescence optical sectioning: Three‐dimensional imaging of macroscopic biological specimens},
  author={Arne H. Voie and David H. Burns and Francis A. Spelman},
  journal={Journal of Microscopy},
An imaging technique called orthogonal‐plane fluorescence optical sectioning (OPFOS) was developed to image the internal architecture of the cochlea. Expressions for the three‐dimensional point spread function and the axial and lateral resolution are derived. Methodologies for tissue preparation and for construction, alignment, calibration and characterization of an OPFOS apparatus are presented. The instrument described produced focused, high‐resolution images of optical sections of an intact… 

Tomographic imaging of macroscopic biomedical objects in high resolution and three dimensions using orthogonal-plane fluorescence optical sectioning.

A new optical-fluorescence microscopy technique, called HR-OPFOS, is discussed and situated among similar OPFOS-implementations, which delivers cross sections through the object under study with semi-histological detail and is demonstrated on gerbil hearing organs and on mouse and bird brains.

Orthogonal-plane fluorescence optical sectioning : a technique for 3-D imaging of biomedical specimens

A popular technique for morphological and structural research is Serial Histological Sectioning. In this method, thin sliced sections of fixed, embedded and stained tissue are created, which are then

Three‐dimensional reconstruction of the guinea pig inner ear, comparison of OPFOS and light microscopy, applications of 3D reconstruction

Three‐dimensional reconstruction of anatomical structures can give additional insight into the morphology and function of these structures and orthogonal plane fluorescence optical sectioning microscopy is a superior technique for 3D reconstruction of inner ear structures in animals.

The OPFOS Microscopy Family: High-Resolution Optical Sectioning of Biomedical Specimens

In this paper, the first implementation of LSFM to image biomedical tissue in three dimensions—orthogonal-plane fluorescence optical sectioning microscopy (OPFOS)—is discussed and the applicability is illustrated on several specimen types with application in biomedical and life sciences.

Light Sheet Fluorescence Microscopy (LSFM)

Light sheet fluorescent microscopy (LSFM), a century‐old idea made possible with modern developments in both excitation and detection optics, provides sub‐cellular resolution and optical sectioning capabilities without compromising speed or excitation efficiency.

Four-dimensional visualization of zebrafish cardiovascular and vessel dynamics by a structured illumination microscope with electrically tunable lens.

Time lapse imaging clearly shows the contractile-relaxation response of the beating heart at different cardiac phases and with different mobilities of blood vessels in different regions in small-sized animals.

Thin-sheet laser imaging microscopy for optical sectioning of thick tissues.

A modular and optimized thin-sheet laser imaging microscope (TSLIM) for nondestructive optical sectioning of organisms and thick tissues such as the mouse cochlea, zebrafish brain/inner ear, and rat brain at a resolution that is comparable to wide-field fluorescence microscopy is reported.

Light Sheet Fluorescence Microscopy

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This review describes development of the technology, reviews existing devices, provides details of one LSFM device, and shows examples of images and three-dimensional reconstructions of tissues that were produced by LSFm.

Light sheet‐based fluorescence microscopy: More dimensions, more photons, and less photodamage

The principles of LSFM are introduced, the challenges of specimen preparation are explained, and the basics of a microscopy that takes advantage of multiple views are introduced.

Light-sheet-based fluorescence microscopy for three-dimensional imaging of biological samples.

The basic principles of LSFM and methods for the preparation, embedding, and imaging of 3D specimens used in the life sciences are discussed in an implementation of L SFM known as the single (or selective) plane illumination microscope (SPIM).



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The improved resolution and sectioning capability of a confocal microscope make it an ideal instrument for extracting three‐dimensional information especially from extended biological specimens and a number of biological applications demonstrated.

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The three-dimensional optical transfer function of a confocal fluorescence microscope is derived, which has no missing cone and provides tomographic images of the sample and the diffraction limitation by an objective lens is verified.

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The advent of relatively inexpensive computers and digital image acquisition systems has now made possible the three-dimensional reconstruction of images taken from the optical microscope.

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A confocal laser microscope scanner for digital recording of optical serial sections

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