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 JoVE Biology

Фазового контраста и дифференциальных помех контраста (DIC) микроскопии

1

1Core Optical Imaging Facility, University of Texas Health Science Center at San Antonio (UTHSCSA)

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    Summary

    Этот протокол освещаются принципы и практические применения фазы и контраста дифференциальных помех (DIC) микроскопии

    Date Published: 8/06/2008, Issue 18; doi: 10.3791/844

    Cite this Article

    Centonze Frohlich, V. Phase Contrast and Differential Interference Contrast (DIC) Microscopy. J. Vis. Exp. (18), e844, doi:10.3791/844 (2008).

    Abstract

    Фазово-контрастной микроскопии часто используется для создания контраста для прозрачной, не поглощающих свет, биологических образцов. Техника была открыта Цернике, в 1942 году, который получил Нобелевскую премию за его достижения. DIC микроскопии, введенный в конце 1960-х, был популярен в биомедицинских исследованиях, поскольку он выделяет края образца структурных деталей, обеспечивает высокое разрешение оптического разделах толстых образцов, включая клетки тканей, яиц и эмбрионов и не страдает от типичных фазы ореолы фазового контраста изображения. Этот протокол освещаются принципы и практическое применение этих методов микроскопии.

    Protocol

    Полный текст протокола для этого экспериментальный подход доступен в Текущий протоколов в клеточной биологии.

    Disclosures

    Acknowledgements

    Несколько клипов видео, демонстрирующее фазового контраста и DIC микроскопии были взяты из другого видео-статью, озаглавленную Слои симбиоз . Юпитер любезно это признает вклад Лидбеттер лаборатории.

    Comments

    9 Comments

    how much will each cost
    Reply

    Posted by: AnonymousSeptember 15, 2008, 1:23 PM

    The systems vary widely to correlate with your needs.  I would be happy to help you further. 631-547-8501, Ned Jastromb, Product Manager, Nikon Instruments, Inc.
    Reply

    Posted by: AnonymousSeptember 16, 2008, 11:15 AM

    Excellent - I'm definitely going to use this for my students, who are from a mixed background (biologists, physicists, chemists, engineers).
    Reply

    Posted by: AnonymousSeptember 18, 2008, 8:40 AM

    I've seen that there we're colored images obtained using the DIC microscopy that looks like a 3-D image. Is there an approximation using DIC to measure the depth or height difference by just interpreting color differences?
    Reply

    Posted by: AnonymousMay 27, 2009, 7:42 AM

    I've seen that there we're colored images obtained using the DIC microscopy that looks like a 3-D image. Is there an approximation using DIC to measure the depth or height difference by just interpreting color differences?
    Reply

    Posted by: AnonymousMay 27, 2009, 7:59 AM

    In article abstract, the phase-contrast microscopy is claimed to have been invented in 194². This is incorrect. The origins of phase-contrast date back to 1934 (not a microscopy yet) and 1935 (microscopy):

    Zernike F. (1934)
    Diffraction theory of the knife-edge test and its improved form, the phase-contrast method.
    Monthly Notices of the Royal Astronomical Society 94 (5), 377-3­84 (originally published in Dutch).
    (http://esoads.eso.org/journals_service.html)

    Zernike F. (1935)
    Phase-contrast method in microscopical observation (in German).
    Physikalische Zeitschrift 36 (²²/²3): 848-851
    Zeitschrift für Technische Physik 16 (11): 454-457
    (two identical papers)

    Furthermore, the following reports (published when Zernike was only 16 or 17 years old) utilize, in a way, the principle of phase-contrast microscopy, even though only for absorbing (i.e., not phase) objects:

    Rheinberg J. (1904 and 1905)
    On the influence on images of gratings of phase differences amongst their spectra.
    J. Roy. Microsc. Soc.; volumes-not-numbered (Part 4), 388-390; and (Part ²), 15²-155.
    (www.biodiversitylibrary.org/bibliography/7413)

    Conrady A. (1905)
    An experimental proof of phase reversal in diffraction spectra.
    J. Roy. Microsc. Soc.; volume-not-numbered (Part ²), 150-15².
    (www.biodiversitylibrary.org/bibliography/7413)
    Reply

    Posted by: AnonymousMay 4, 2010, 11:14 AM

    Your presentation is excellent and locked in my neurons , the corresponding text in books do not give that effect
    Reply

    Posted by: AnonymousNovember 12, 2011, 10:19 PM

    Nice video.
    I may be mistaken, but one of the images (around 3:47) shown as DIC appears to be phase contrast.
    Also, when mentioning setting up KŒhler illumination during DIC (around 4:30), the video shows the alignment of phase rings. This is not needed (since you do not need to see the back focal plane of the microscope to establish KŒhler) nor, in fact, what would be observed when setting up KŒhler for DIC (there would be no phase annulus in the condenser if it is set up for DIC).
    Reply

    Posted by: AnonymousDecember 7, 2011, 10:36 AM

    The video explained well what i was trying to learn in detail....
    it is really nice
    Thanks
    Reply

    Posted by: AnonymousApril 5, 2012, 2:35 AM

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