Dual-detection confocal microscopy: high-speed surface profiling without depth scanning

2016 ◽  
Author(s):  
Dong-Ryoung Lee ◽  
Dae-Gab Gweon ◽  
Hongki Yoo
Keyword(s):  
Confocal Microscopy ◽  
High Speed ◽  
Surface Profiling ◽  
Dual Detection

scholarly journals Swept-Source-Based Chromatic Confocal Microscopy

Sensors ◽  
2020 ◽  
Vol 20 (24) ◽  
pp. 7347
Author(s):  
Dawoon Jeong ◽  
Se Jin Park ◽  
Hansol Jang ◽  
Hyunjoo Kim ◽  
Jaesun Kim ◽  
...  
Keyword(s):  
Confocal Microscopy ◽  
High Speed ◽  
Imaging System ◽  
Three Dimensional ◽  
Chromatic Aberration ◽  
Cover Glass ◽  
Focal Distance ◽  
Swept Source ◽  
Surface Profiling ◽  
Novel Concept

Chromatic confocal microscopy (CCM) has been intensively developed because it can exhibit effective focal position scanning based on the axial chromatic aberration of broadband light reflected from a target. To improve the imaging speed of three-dimensional (3D) surface profiling, we have proposed the novel concept of swept-source-based CCM (SS-CCM) and investigated the usefulness of the corresponding imaging system. Compared to conventional CCM based on a broadband light source and a spectrometer, a swept-source in the proposed SS-CCM generates light with a narrower linewidth for higher intensity, and a single photodetector employed in the system exhibits a fast and sensitive response by immediately obtaining spectrally encoded depth from a chromatic dispersive lens array. Results of the experiments conducted to test the proposed SS-CCM system indicate that the system exhibits an axial chromatic focal distance range of approximately 360 μm for the 770–820 nm swept wavelength range. Moreover, high-speed surface profiling images of a cover glass and coin were successfully obtained with a short measurement time of 5 ms at a single position.

Five-Dimensional Microscopy using Widefield-Deconvolution: Practical Considerations and Biological Applications

1996 ◽  
Vol 54 ◽  
pp. 268-269
Author(s):  
W.F. Marshall ◽  
K. Oegema ◽  
J. Nunnari ◽  
A.F. Straight ◽  
D.A. Agard ◽  
...  
Keyword(s):  
Confocal Microscopy ◽  
High Speed ◽  
Laser Scanning ◽  
Ccd Camera ◽  
Image Plane ◽  
Time Lapse ◽  
Laser Scanning Confocal Microscopy ◽  
Three Dimensions ◽  
Excitation Light ◽  
Cooled Ccd

The ability to image cells in three dimensions has brought about a revolution in biological microscopy, enabling many questions to be asked which would be inaccessible without this capability. There are currently two major methods of three dimensional microscopy: laser-scanning confocal microscopy and widefield-deconvolution microscopy. The method of widefield-deconvolution uses a cooled CCD to acquire images from a standard widefield microscope, and then computationally removes out of focus blur. Using such a scheme, it is easy to acquire time-lapse 3D images of living cells without killing them, and to do so for multiple wavelengths (using computer-controlled filter wheels). Thus, it is now not only feasible, but routine, to perform five dimensional microscopy (three spatial dimensions, plus time, plus wavelength).Widefield-deconvolution has several advantages over confocal microscopy. The two main advantages are high speed of acquisition (because there is no scanning, a single optical section is acquired at a time by using a cooled CCD camera) and the use of low excitation light levels Excitation intensity can be much lower than in a confocal microscope for three reasons: 1) longer exposures can be taken since the entire 512x512 image plane is acquired in parallel, so that dwell time is not an issue, 2) the higher quantum efficiently of a CCD detect over those typically used in confocal microscopy (although this is expected to change due to advances in confocal detector technology), and 3) because no pinhole is used to reject light, a much larger fraction of the emitted light is collected. Thus we can typically acquire images with thousands of photons per pixel using a mercury lamp, instead of a laser, for illumination. The use of low excitation light is critical for living samples, and also reduces bleaching. The high speed of widefield microscopy is also essential for time-lapse 3D microscopy, since one must acquire images quickly enough to resolve interesting events.

Simultaneous multiplane imaging for 3D confocal microscopy using high-speed z-scanning multiplexing

2015 ◽  
Author(s):  
Marti Duocastella ◽  
Giuseppe Vicidomini ◽  
Alberto Diaspro
Keyword(s):  
Confocal Microscopy ◽  
High Speed
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