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Contraste de fase e de interferência diferencial Contrast Microscopy (DIC)

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

 

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Cite this Article: Contraste de fase e de interferência diferencial Contrast Microscopy (DIC)

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

Abstract: Contraste de fase e de interferência diferencial Contrast Microscopy (DIC)

Microscopia de contraste de fase é muitas vezes usado para produzir contraste para transparente, não de absorção de luz, as amostras biológicas. A técnica foi descoberta por Zernike, em 1942, que recebeu o prêmio Nobel por suas realizações. Microscopia DIC, introduzido no final dos anos 1960, tem sido popular na pesquisa biomédica, porque enfatiza bordas do espécime detalhes estruturais, proporciona alta resolução secções ópticas de amostras de espessura, incluindo as células dos tecidos, ovos e embriões e não sofrem com o halos fase típica de contraste de fase imagens. Este protocolo destaca os princípios e aplicações práticas dessas técnicas de microscopia.

Protocol: Contraste de fase e de interferência diferencial Contrast Microscopy (DIC)

O protocolo de texto completo para essa abordagem experimental está disponível em protocolos atuais em biologia celular.

Disclosures: Contraste de fase e de interferência diferencial Contrast Microscopy (DIC)

Acknowledgements: Contraste de fase e de interferência diferencial Contrast Microscopy (DIC)

Vários clips de vídeo demonstrando contraste de fase e microscopia DIC foram tiradas de um outro vídeo-artigo, intitulado Camadas de simbiose . JOVE graciosamente reconhece esta contribuição feita pelo laboratório Leadbetter.

Ask the Author: Contraste de fase e de interferência diferencial Contrast Microscopy (DIC)

8 Comments

how much will each cost

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Posted by: abrahamSeptember 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.

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Posted by: jastrombSeptember 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).

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Posted by: Dr. Una FitzGeraldSeptember 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?

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Posted by: KisutoMay 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?

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Posted by: KisutoMay 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)

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Posted by: Radek PelcMay 4, 2010, 11:14 AM

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

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Posted by: Brajendra Kumar SinhaNovember 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 Koehler 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 Koehler) nor, in fact, what would be observed when setting up Koehler for DIC (there would be no phase annulus in the condenser if it is set up for DIC).

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Posted by: Eric RobertsDecember 7, 2011, 10:36 AM

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

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

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