Differential high-speed digital micromirror device based fluorescence speckle confocal microscopy

2010 ◽  
Vol 49 (3) ◽  
pp. 497 ◽  
Author(s):  
Shihong Jiang ◽  
John Walker
Author(s):  
W.F. Marshall ◽  
K. Oegema ◽  
J. Nunnari ◽  
A.F. Straight ◽  
D.A. Agard ◽  
...  

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.


2021 ◽  
Vol 19 (9) ◽  
pp. 093301
Author(s):  
Kohei Suzuki ◽  
Minori Tao ◽  
Yuki Maeda ◽  
Hirotaka Nakayama ◽  
Ren Noguchi ◽  
...  

2013 ◽  
Vol 50 (10) ◽  
pp. 101702
Author(s):  
涂龙 Tu Long ◽  
余锦 Yu Jin ◽  
樊仲维 Fan Zhongwei ◽  
邱基斯 Qiu Jisi ◽  
赵天卓 Zhao Tianzhuo ◽  
...  

Optik ◽  
2019 ◽  
Vol 185 ◽  
pp. 1036-1044 ◽  
Author(s):  
Chao Peng ◽  
Zezhou Zhang ◽  
Jianxiao Zou ◽  
Wenming Chi

2015 ◽  
Author(s):  
Marti Duocastella ◽  
Giuseppe Vicidomini ◽  
Alberto Diaspro

2015 ◽  
Vol 40 (21) ◽  
pp. 4875 ◽  
Author(s):  
Jiyi Cheng ◽  
Chenglin Gu ◽  
Dapeng Zhang ◽  
Shih-Chi Chen

2014 ◽  
Author(s):  
Tien-Hsin Chao ◽  
Thomas Lu ◽  
Brian Walker ◽  
George Reyes

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