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Contraste de phase et les interférences de contraste différentiel (DIC) Microscopie

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

 

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Cite this Article: Contraste de phase et les interférences de contraste différentiel (DIC) Microscopie

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

Abstract: Contraste de phase et les interférences de contraste différentiel (DIC) Microscopie

Phase microscopie à contraste est souvent utilisé pour produire de contraste pour transparentes, non absorbant la lumière, des spécimens biologiques. La technique a été découvert par Zernike, en 1942, qui a reçu le prix Nobel pour sa réalisation. Microscopie DIC, introduit dans la fin des années 1960, a été populaire dans la recherche biomédicale, car il souligne les bords de détail spécimen structurelles, offre une haute résolution de sections optiques des échantillons d'épaisseur, y compris les cellules des tissus, des œufs et des embryons et ne souffre pas de halos de phase typique des images à contraste de phase. Ce protocole met en évidence les principes et les applications pratiques de ces techniques de microscopie.

Protocol: Contraste de phase et les interférences de contraste différentiel (DIC) Microscopie

Le protocole texte complet de cette approche expérimentale est disponible en Current Protocols in Cell Biology.

Disclosures: Contraste de phase et les interférences de contraste différentiel (DIC) Microscopie

Acknowledgements: Contraste de phase et les interférences de contraste différentiel (DIC) Microscopie

Plusieurs clips vidéo montrant le contraste de phase et la microscopie DIC ont été prélevés sur une autre vidéo-article, intitulé Des couches de Symbiose . JoVE reconnaît gracieusement cette contribution faite par le laboratoire de Leadbetter.

Ask the Author: Contraste de phase et les interférences de contraste différentiel (DIC) Microscopie

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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