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Microscopy
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==== Wide-field multiphoton microscopy ==== {{Main|Wide-field multiphoton microscopy}}Wide-field multiphoton microscopy<ref name=":1">{{Cite journal|last1=Peterson|first1=Mark D.|last2=Hayes|first2=Patrick L.|last3=Martinez|first3=Imee Su|last4=Cass|first4=Laura C.|last5=Achtyl|first5=Jennifer L.|last6=Weiss|first6=Emily A.|last7=Geiger|first7=Franz M.|date=2011-05-01|title=Second harmonic generation imaging with a kHz amplifier [Invited]|journal=Optical Materials Express|language=EN|volume=1|issue=1|doi=10.1364/ome.1.000057|page=57|bibcode=2011OMExp...1...57P}}</ref><ref name=":0">{{Cite journal|last1=Macias-Romero|first1=Carlos|last2=Didier|first2=Marie E. P.|last3=Jourdain|first3=Pascal|last4=Marquet|first4=Pierre|last5=Magistretti|first5=Pierre|last6=Tarun|first6=Orly B.|last7=Zubkovs|first7=Vitalijs|author8-link=Aleksandra Radenovic|last8=Radenovic|first8=Aleksandra|last9=Roke|first9=Sylvie|date=2014-12-15|title=High throughput second harmonic imaging for label-free biological applications|journal=Optics Express|language=EN|volume=22|issue=25|pages=31102β12|doi=10.1364/oe.22.031102|pmid=25607059|bibcode=2014OExpr..2231102M|url=http://infoscience.epfl.ch/record/203737|doi-access=free|access-date=2019-12-11|archive-date=2020-06-20|archive-url=https://web.archive.org/web/20200620070847/https://infoscience.epfl.ch/record/203737|url-status=live|hdl=10754/563269|hdl-access=free}}</ref><ref name=":2">{{Cite journal|last1=Cheng|first1=Li-Chung|last2=Chang|first2=Chia-Yuan|last3=Lin|first3=Chun-Yu|last4=Cho|first4=Keng-Chi|last5=Yen|first5=Wei-Chung|last6=Chang|first6=Nan-Shan|last7=Xu|first7=Chris|last8=Dong|first8=Chen Yuan|last9=Chen|first9=Shean-Jen|date=2012-04-09|title=Spatiotemporal focusing-based widefield multiphoton microscopy for fast optical sectioning|journal=Optics Express|language=EN|volume=20|issue=8|pages=8939β48|doi=10.1364/oe.20.008939|pmid=22513605|bibcode=2012OExpr..20.8939C|doi-access=free}}</ref><ref name=":3">{{Cite journal|last1=Oron|first1=Dan|last2=Tal|first2=Eran|last3=Silberberg|first3=Yaron|date=2005-03-07|title=Scanningless depth-resolved microscopy|journal=Optics Express|language=EN|volume=13|issue=5|pages=1468β76|doi=10.1364/opex.13.001468|pmid=19495022|bibcode=2005OExpr..13.1468O|doi-access=free}}</ref> refers to an [[Nonlinear optics|optical non-linear imaging technique]] in which a large area of the object is illuminated and imaged without the need for scanning. High intensities are required to induce non-linear optical processes such as [[Two-photon excitation microscopy|two-photon fluorescence]] or [[Second-harmonic generation|second harmonic generation]]. In [[Multiphoton fluorescence microscope|scanning multiphoton microscopes]] the high intensities are achieved by tightly focusing the light, and the image is obtained by beam scanning. In '''wide-field multiphoton microscopy''' the high intensities are best achieved using an [[Optical amplifier|optically amplified]] pulsed laser source to attain a large field of view (~100 ΞΌm).<ref name=":1" /><ref name=":0" /><ref name=":2" /> The image in this case is obtained as a single frame with a CCD camera without the need of scanning, making the technique particularly useful to visualize dynamic processes simultaneously across the object of interest. With wide-field multiphoton microscopy the frame rate can be increased up to a 1000-fold compared to [[Multiphoton fluorescence microscopy|multiphoton scanning microscopy]].<ref name=":0" /> In scattering tissue, however, image quality rapidly degrades with increasing depth.
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