Three-dimensional quantification with laser scanning confocal microscopy

Takahiro Ode; Satoshi Komiya

doi:10.1117/12.165183

29 December 1993 Three-dimensional quantification with laser scanning confocal microscopy

Takahiro Ode, Satoshi Komiya

Proceedings Volume 1987, Recording Systems: High-Resolution Cameras and Recording Devices and Laser Scanning and Recording Systems; (1993) https://doi.org/10.1117/12.165183
Event: Electronic Imaging Device Engineering, 1993, Munich, Germany

Abstract

A confocal microscope system illuminates the specimen with a point light source and then collects the reflected light onto a point detector. Only when the specimen is exactly in focus will any significant amount of light fall onto the detector. This gives the height sectioning capability of the microscope. Obtaining 0.1 micron depth resolution is not difficult. A potential drawback of the confocal microscope is that only the in focus region of the specimen is visible. We can use this property to form both an extended focus image and a three dimensional topological map. This extended focus mode is useful for understanding three dimensional structures. By using a three dimensional topological map image, where each pixel has an accurate calibration in three dimension, one can calculate various parameters such as surface roughness, height, depth, and volume. The volume of a three dimensional structure can be defined as the integral of the individual pixel values lying under the reference plane. One can define this reference plane manually with a mouse and a screen cursor. The software finds the volume below the surface enclosed within that area, deriving the height of the surrounding, reference surface.

Citation Download Citation

Takahiro Ode and Satoshi Komiya "Three-dimensional quantification with laser scanning confocal microscopy", Proc. SPIE 1987, Recording Systems: High-Resolution Cameras and Recording Devices and Laser Scanning and Recording Systems, (29 December 1993); https://doi.org/10.1117/12.165183

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available