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High-speed microscopy of continuously moving cell culture vessels

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ABSTRACT

We report a method of high-speed phase contrast and bright field microscopy which permits large cell culture vessels to be scanned at much higher speed (up to 30 times faster) than when conventional methods are used without compromising image quality. The object under investigation moves continuously and is captured using a flash illumination which creates an exposure time short enough to prevent motion blur. During the scan the object always stays in focus due to a novel hardware-autofocus system.

No MeSH data available.


Stitched image of continuous imaging scan of one well of a 6-well microtiter plate filled with adherent iPS cell colonies on Matrigel cultivated in mTeSR.Magnification: 4x, imaging mode: phase contrast, scan velocity: 34 mm/s, pre-processing: shading correction.
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f7: Stitched image of continuous imaging scan of one well of a 6-well microtiter plate filled with adherent iPS cell colonies on Matrigel cultivated in mTeSR.Magnification: 4x, imaging mode: phase contrast, scan velocity: 34 mm/s, pre-processing: shading correction.

Mentions: Besides creating single images with uncompromised quality that are free of motion blur due to the short flash exposure and adequately focused by virtue of the novel hardware autofocus system, our solution provides excellent image mosaics to display large parts of the sample at high resolution (Fig. 7). We achieve this by using a fast stitching approach (Methods section). Other preprocessing steps to enhance the image quality include a camera calibration, shading correction and histogram normalization.


High-speed microscopy of continuously moving cell culture vessels
Stitched image of continuous imaging scan of one well of a 6-well microtiter plate filled with adherent iPS cell colonies on Matrigel cultivated in mTeSR.Magnification: 4x, imaging mode: phase contrast, scan velocity: 34 mm/s, pre-processing: shading correction.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5036042&req=5

f7: Stitched image of continuous imaging scan of one well of a 6-well microtiter plate filled with adherent iPS cell colonies on Matrigel cultivated in mTeSR.Magnification: 4x, imaging mode: phase contrast, scan velocity: 34 mm/s, pre-processing: shading correction.
Mentions: Besides creating single images with uncompromised quality that are free of motion blur due to the short flash exposure and adequately focused by virtue of the novel hardware autofocus system, our solution provides excellent image mosaics to display large parts of the sample at high resolution (Fig. 7). We achieve this by using a fast stitching approach (Methods section). Other preprocessing steps to enhance the image quality include a camera calibration, shading correction and histogram normalization.

View Article: PubMed Central - PubMed

ABSTRACT

We report a method of high-speed phase contrast and bright field microscopy which permits large cell culture vessels to be scanned at much higher speed (up to 30 times faster) than when conventional methods are used without compromising image quality. The object under investigation moves continuously and is captured using a flash illumination which creates an exposure time short enough to prevent motion blur. During the scan the object always stays in focus due to a novel hardware-autofocus system.

No MeSH data available.