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High-speed microscopy of continuously moving cell culture vessels

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.


Spectrum (a-scan) of the focus measurement after applying a fast Fourier transform (FFT) to the interference raw signal.The lower (p1) and upper (p2) reflection peaks from the microtiter plate bottom are clearly visible. There is another peak formed through the autocorrelation of the two plate bottom reflections which resides around pixel position 390.
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f5: Spectrum (a-scan) of the focus measurement after applying a fast Fourier transform (FFT) to the interference raw signal.The lower (p1) and upper (p2) reflection peaks from the microtiter plate bottom are clearly visible. There is another peak formed through the autocorrelation of the two plate bottom reflections which resides around pixel position 390.

Mentions: In contrast to commercially available laser-based focus measurement systems, our system is able to resolve multiple interfaces of the measured object simultaneously. Each reflection from an interface of the sample leads to a peak in the spectrum (Fig. 5, peak 1 = plate underside, interface air-plastic, peak 2 = plate upper side, interface plastic-medium) that is determined at subpixel resolution with a peak fit algorithm (Methods section).


High-speed microscopy of continuously moving cell culture vessels
Spectrum (a-scan) of the focus measurement after applying a fast Fourier transform (FFT) to the interference raw signal.The lower (p1) and upper (p2) reflection peaks from the microtiter plate bottom are clearly visible. There is another peak formed through the autocorrelation of the two plate bottom reflections which resides around pixel position 390.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f5: Spectrum (a-scan) of the focus measurement after applying a fast Fourier transform (FFT) to the interference raw signal.The lower (p1) and upper (p2) reflection peaks from the microtiter plate bottom are clearly visible. There is another peak formed through the autocorrelation of the two plate bottom reflections which resides around pixel position 390.
Mentions: In contrast to commercially available laser-based focus measurement systems, our system is able to resolve multiple interfaces of the measured object simultaneously. Each reflection from an interface of the sample leads to a peak in the spectrum (Fig. 5, peak 1 = plate underside, interface air-plastic, peak 2 = plate upper side, interface plastic-medium) that is determined at subpixel resolution with a peak fit algorithm (Methods section).

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.