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{{Infobox laboratory equipment|frame
{{Infobox laboratory equipment|frame
|name = Quantitative phase contrast microscope
|name = Quantitative phase contrast microscope
|image = HoloMonitor M4 with IBIDI slide.png
|image =
|alt = <!-- Wikipedia:Alternative text for images -->
|alt = <!-- Wikipedia:Alternative text for images -->
|caption = A quantitative phase contrast microscope imaging cultured cells in a slide based cell culture vessel.
|caption = A quantitative phase contrast microscope imaging cultured cells in a cell culture incubator.
|acronym = QPCM
|acronym = QPCM, QPM, QPI
|other_names = Phase microscope, Quantitative phase microscopy
|other_names = Phase microscope, Quantitative phase microscopy, Quantitative phase imaging
|uses = Microscopic observation and quantification of unstained biological material
|uses = Microscopic observation and quantification of unstained biological material
|notable_experiments =
|notable_experiments =
⚫
|related = [[Phase contrast microscopy]], [[Differential interference contrast microscopy]], [[Hoffman modulation contrast|Hoffman modulation contrast microscopy]]
|manufacturer =
|model =
⚫
|related = [[Phase contrast microscopy]], [[Differential interference contrast microscopy]], [[Hoffman modulation contrast|Hoffman modulation contrast microscopy]], [[Ptychography]]
}}
}}
{{Advert|date=October 2018}}

'''Quantitative phase contrast microscopy''' is the collective name for a group of microscopy methods that quantify the [[Phase (waves)|phase shift]] that occurs when light waves pass through a more optically dense object.
'''Quantitative phase contrast microscopy''' or '''quantitative phase imaging''' are the collective names for a group of microscopy methods that quantify the [[Phase (waves)|phase shift]] that occurs when light waves pass through a more optically dense object.<ref name="Cuche">{{cite journal

⚫
Translucent objects, like a living human cell, absorb and scatter small amounts of light.
⚫
This makes translucent objects difficult to observe in ordinary light microscopes.
⚫
Such objects do, however, induce a phase shift that can be observed using a [[phase contrast microscopy|phase contrast microscope]].
⚫
Conventional phase contrast microscopy and related methods, such as [[differential interference contrast microscopy]], quantitative phase microscopy, or digital holographic microscopy, visualize phase shifts by transforming phase shift gradients into intensity variations.
⚫
These intensity variations are mixed with other intensity variations, making it difficult to extract quantitative information.<ref name="Cuche">{{cite journal
| author=Etienne Cuche |author2=Frédéric Bevilacqua |author3=Christian Depeursinge
| author=Etienne Cuche |author2=Frédéric Bevilacqua |author3=Christian Depeursinge
| title=Digital holography for quantitative phase-contrast imaging
| title=Digital holography for quantitative phase-contrast imaging
Line 28: Line 20:
| pages=291–293
| pages=291–293
| year=1999
| year=1999
| doi=10.1364/OL.24.000291|bibcode = 1999OptL...24..291C }}</ref><ref name="Marquet">{{cite journal
| doi=10.1364/OL.24.000291|pmid=18071483 |bibcode = 1999OptL...24..291C }}</ref><ref>{{cite journal
| vauthors=Park Y, Depeursinge C, [[Gabriel Popescu (scientist)|Popescu G]]
| author=Pierre Marquet |author2=Benjamin Rappaz |author3=Pierre J. Magistretti |author4=Etienne Cuche |author5=Yves Emery |author6=Tristan Colomb |author7=Christian Depeursinge
| title=Quantitative phase imaging in biomedicine
| title=Digital holographic microscopy: a noninvasive contrast imaging technique allowing quantitative visualization of living cells with subwavelength axial accuracy
| journal=Optics Letters
| journal=Nature Photonics
| volume=30
| date=2018
| issue=5
| volume=12
⚫
| issue=10
| pages=468–470
| pages=578–589
⚫
| year=2005
| doi=10.1038/s41566-018-0253-x| pmid=<!--none-->
| doi=10.1364/OL.30.000468|bibcode = 2005OptL...30..468M }}</ref><ref>{{Cite book|url=https://www.worldcat.org/oclc/659763966|title=Quantitative phase imaging of cells and tissues|last=1971-|first=Popescu, Gabriel,|date=2011-01-01|publisher=McGraw-Hill|isbn=0071663428|oclc=659763966}}</ref><ref>{{Cite journal|last=Lee|first=KyeoReh|last2=Kim|first2=Kyoohyun|last3=Jung|first3=Jaehwang|last4=Heo|first4=JiHan|last5=Cho|first5=Sangyeon|last6=Lee|first6=Sangyun|last7=Chang|first7=Gyuyoung|last8=Jo|first8=YoungJu|last9=Park|first9=Hyunjoo|date=2013-03-28|title=Quantitative Phase Imaging Techniques for the Study of Cell Pathophysiology: From Principles to Applications|url=http://www.mdpi.com/1424-8220/13/4/4170|journal=Sensors|language=en|volume=13|issue=4|pages=4170–4191|doi=10.3390/s130404170|pmc=3673078|pmid=23539026}}</ref>
| bibcode=2018NaPho..12..578P
| s2cid=256704142
}}</ref>

⚫
Translucent objects, like a living human cell, absorb and scatter small amounts of light.
⚫
This makes translucent objects much easier to observe in ordinary light microscopes.
⚫
Such objects do, however, induce a phase shift that can be observed using a [[phase contrast microscopy|phase contrast microscope]].
⚫
Conventional phase contrast microscopy and related methods, such as [[differential interference contrast microscopy]], visualize phase shifts by transforming phase shift gradients into intensity variations.
⚫
These intensity variations are mixed with other intensity variations, making it difficult to extract quantitative information.


Quantitative phase contrast methods are distinguished from other phase contrast methods in that
Quantitative phase contrast methods are distinguished from conventional phase contrast methods in that
they create a second so-called ''phase shift image'' or ''phase image'', independent of the intensity ([[Bright field microscopy|bright field]]) image.
they create a second so-called ''phase shift image'' or ''phase image'', independent of the intensity ([[Bright field microscopy|bright field]]) image.
[[Phase unwrapping]] methods are generally applied to the phase shift image to give absolute phase shift
[[Phase unwrapping]] methods are generally applied to the phase shift image to give absolute phase shift
values in each pixel, as exemplified by Figure 1.
values in each pixel, as exemplified by Figure 1.


⚫
[[File:Phase shift image of cells in 3D.jpg|thumb| '''Figure 1''': In this phase shift image of cells in culture, the height and color of an image point correspond to the measured phase shift. The phase shift induced by an object in an image point depends only on the object thickness and the relative [[refractive index]] of the object in the image point. The volume of an object can therefore be determined from a phase shift image when the difference in refractive index between the object and the surrounding media is known.<ref name="Kemmler">{{cite journal
The principal methods for measuring and visualizing phase shifts include [[ptychography]],<ref>{{cite journal|vauthors = Marrison J, Räty L, Marriott P, O'Toole P |title=Ptychography – a label free, high-contrast imaging technique for live cells using quantitative phase information|journal=Scientific Reports|date=6 August 2013|volume=3|issue=2369|doi=10.1038/srep02369|url=http://www.nature.com/srep/2013/130806/srep02369/full/srep02369.html|bibcode = 2013NatSR...3E2369M|pmid=23917865|pmc=3734479}}</ref> [[Fourier ptychography]],<ref>{{cite journal|last1=Zheng|first1=Guoan|last2=Horstmeyer|first2=Roarke|last3=Yang|first3=Changhuei|title=Wide-field, high-resolution Fourier ptychographic microscopy|journal=Nature Photonics|date=29 July 2013|volume=7|doi=10.1038/nphoton.2013.187|url=http://www.nature.com/nphoton/journal/v7/n9/full/nphoton.2013.187.html|pages=739–745}}</ref> and various types
⚫
| author=Manuel Kemmler |author2=Markus Fratz |author3=Dominik Giel |author4=Norbert Saum |author5=Albrecht Brandenburg |author6=Christian Hoffmann
⚫
| title=Noninvasive time-dependent cytometry monitoring by digital holography
⚫
| journal=Journal of Biomedical Optics
⚫
| volume=12
⚫
| issue=6
⚫
| pages=064002
⚫
| year=2007
⚫
| doi=10.1117/1.2804926
⚫
| pmid=18163818|bibcode = 2007JBO....12f4002K |s2cid=40335328 | doi-access=free}}</ref>]]

The principal methods for measuring and visualizing phase shifts include [[ptychography]] and various types
of holographic microscopy methods such as [[digital holographic microscopy]],
of holographic microscopy methods such as [[digital holographic microscopy]],
[[holographic interference microscopy]] and digital in-line holographic microscopy.<ref name="4Deep inwater imaging">{{cite web
[[holographic interference microscopy]] and digital in-line holographic microscopy.
| title=4Deep inwater imaging
| url=http://www.4-deep.com}}</ref>
Common to these methods is that an [[Interference (wave propagation)|interference pattern]] ([[hologram]]) is recorded by a digital [[image sensor]].
Common to these methods is that an [[Interference (wave propagation)|interference pattern]] ([[hologram]]) is recorded by a digital [[image sensor]].
From the recorded interference pattern, the intensity and the phase shift image is numerically created by a computer [[algorithm]].<ref name="Kim">{{cite journal
From the recorded interference pattern, the intensity and the phase shift image is numerically created by a computer [[algorithm]].<ref name="Kim">{{cite journal
Line 57: Line 67:
| year=2010
| year=2010
| doi=10.1117/6.0000006
| doi=10.1117/6.0000006
|bibcode = 2010SPIER...1a8005K }}</ref>
|bibcode = 2010SPIER...1a8005K | doi-access=free
}}</ref>


Conventional phase contrast microscopy is primarily used to observed unstained living cells.<ref>{{cite web
Quantitative phase contrast microscopy is primarily used to observed unstained living cells.
Measuring the phase delay images of biological cells provides quantitative information about the morphology and the drymass of individual cells.<ref>{{cite journal |vauthors=Zangle T, Teitell M |title=Live-cell mass profiling: an emerging approach in quantitative biophysics |journal=Nature Methods |date=2014 |volume=11 |issue=12 |pages=1221–1228 |doi=10.1038/nmeth.3175|pmid=25423019 |pmc=4319180 }}</ref>
| title=The phase-contrast microscope
⚫
Contrary to conventional phase contrast images{{fact|date=October 2020}}, phase shift images of living cells are suitable to be processed by image analysis software.
| publisher=Nobel Media AB
⚫
This has led to the development of non-invasive live cell imaging and automated [[cell culture]] analysis systems based on quantitative phase contrast microscopy.<ref name=CChen>{{cite journal
| url=https://www.nobelprize.org/educational/physics/microscopes/phase}}</ref>
⚫
Measuring the phase delay images of biological cells provides quantitative information about the morphology and the drymass of individual cells. Contrary to conventional phase contrast images, phase shift images of living cells are suitable to be processed by image analysis software.
⚫
This has led to the development of non-invasive live cell imaging and automated [[cell culture]] analysis systems based on quantitative phase contrast microscopy.<ref name="4Deep inwater imaging"/><ref name="Popescu">{{Cite journal
| doi = 10.1364/OL.31.000775
| last1 = Popescu | first1 = G.
| last2 = Ikeda | first2 = T.
| last3 = Dasari | first3 = R. R.
| last4 = Feld | first4 = M. S.
| title = Diffraction phase microscopy for quantifying cell structure and dynamics
| journal = Optics Letters
| volume = 31
⚫
| issue = 6
| pages = 775–777
| year = 2006
| pmid = 16544620
|bibcode = 2006OptL...31..775P }}</ref><ref name="PHI">{{cite web
| title=Quantitative phase contrast imaging microscopy - label-free live cell imaging and analysis
| publisher=Phase Holographic Imaging AB
| url=http://www.phiab.se/technology/quantitative-phase-contrast-microscopy}}</ref><ref>{{cite web
| title=Ovizio Imaging Systems
| url=http://www.ovizio.com}}</ref><ref name=CChen>{{cite journal
| title =Deep Learning in Label-free Cell Classification
| title =Deep Learning in Label-free Cell Classification
| journal =Scientific Reports
| journal =Scientific Reports
Line 88: Line 79:
| issue =
| issue =
| pages =21471
| pages =21471
| publisher =
| location =Full text download available
| date =
| doi =10.1038/srep21471
| doi =10.1038/srep21471
| pmid =26975219
| pmid =26975219
Line 110: Line 98:
| last7 =Jalali
| last7 =Jalali
| first7 =Bahram
| first7 =Bahram
| bibcode =2016NatSR...621471C}}[http://www.nature.com/srep/about/open-access published under CC BY 4.0 licensing]</ref><ref>{{Cite web|url=http://www.tomocube.com/|title=Tomocube, Inc. – The new era of microscopy – 3D Holographic Microscopy|website=www.tomocube.com|access-date=2016-12-30}}</ref>
| bibcode =2016NatSR...621471C}}[http://www.nature.com/srep/about/open-access published under CC BY 4.0 licensing]</ref>

⚫
[[File:Phase shift image of cells in 3D.jpg|thumb| '''Figure 1''': In this phase shift image of cells in culture, the height and color of an image point correspond to the measured phase shift. The phase shift induced by an object in an image point depends only on the object's thickness and the relative [[refractive index]] of the object in the image point. The volume of an object can therefore be determined from a phase shift image when the difference in refractive index between the object and the surrounding media is known.<ref name="Kemmler">{{cite journal
⚫
| author=Manuel Kemmler |author2=Markus Fratz |author3=Dominik Giel |author4=Norbert Saum |author5=Albrecht Brandenburg |author6=Christian Hoffmann
⚫
| title=Noninvasive time-dependent cytometry monitoring by digital holography
⚫
| journal=J. of Biomedical Optics
⚫
| volume=12
⚫
| issue=6
⚫
| pages=064002
| year=2007
⚫
| doi=10.1117/1.2804926
⚫
| pmid=18163818|bibcode = 2007JBO....12f4002K }}</ref>]]


==See also==
==See also==
Line 147: Line 124:
[[Category:Microbiology techniques]]
[[Category:Microbiology techniques]]
[[Category:Laboratory techniques]]
[[Category:Laboratory techniques]]
[[Category:Laboratory equipment]]
[[Category:Optical microscopy]]

Latest revision as of 17:45, 4 August 2024

Quantitative phase contrast microscope
AcronymQPCM, QPM, QPI
Other namesPhase microscope, Quantitative phase microscopy, Quantitative phase imaging
UsesMicroscopic observation and quantification of unstained biological material
Related itemsPhase contrast microscopy, Differential interference contrast microscopy, Hoffman modulation contrast microscopy

Quantitative phase contrast microscopy or quantitative phase imaging are the collective names for a group of microscopy methods that quantify the phase shift that occurs when light waves pass through a more optically dense object.[1][2]

Translucent objects, like a living human cell, absorb and scatter small amounts of light. This makes translucent objects much easier to observe in ordinary light microscopes. Such objects do, however, induce a phase shift that can be observed using a phase contrast microscope. Conventional phase contrast microscopy and related methods, such as differential interference contrast microscopy, visualize phase shifts by transforming phase shift gradients into intensity variations. These intensity variations are mixed with other intensity variations, making it difficult to extract quantitative information.

Quantitative phase contrast methods are distinguished from conventional phase contrast methods in that they create a second so-called phase shift image or phase image, independent of the intensity (bright field) image. Phase unwrapping methods are generally applied to the phase shift image to give absolute phase shift values in each pixel, as exemplified by Figure 1.

Figure 1: In this phase shift image of cells in culture, the height and color of an image point correspond to the measured phase shift. The phase shift induced by an object in an image point depends only on the object thickness and the relative refractive index of the object in the image point. The volume of an object can therefore be determined from a phase shift image when the difference in refractive index between the object and the surrounding media is known.[3]

The principal methods for measuring and visualizing phase shifts include ptychography and various types of holographic microscopy methods such as digital holographic microscopy, holographic interference microscopy and digital in-line holographic microscopy. Common to these methods is that an interference pattern (hologram) is recorded by a digital image sensor. From the recorded interference pattern, the intensity and the phase shift image is numerically created by a computer algorithm.[4]

Quantitative phase contrast microscopy is primarily used to observed unstained living cells. Measuring the phase delay images of biological cells provides quantitative information about the morphology and the drymass of individual cells.[5] Contrary to conventional phase contrast images[citation needed], phase shift images of living cells are suitable to be processed by image analysis software. This has led to the development of non-invasive live cell imaging and automated cell culture analysis systems based on quantitative phase contrast microscopy.[6]

See also

[edit]

References

[edit]
  1. ^ Etienne Cuche; Frédéric Bevilacqua; Christian Depeursinge (1999). "Digital holography for quantitative phase-contrast imaging". Optics Letters. 24 (5): 291–293. Bibcode:1999OptL...24..291C. doi:10.1364/OL.24.000291. PMID 18071483.
  2. ^ Park Y, Depeursinge C, Popescu G (2018). "Quantitative phase imaging in biomedicine". Nature Photonics. 12 (10): 578–589. Bibcode:2018NaPho..12..578P. doi:10.1038/s41566-018-0253-x. S2CID 256704142.
  3. ^ Manuel Kemmler; Markus Fratz; Dominik Giel; Norbert Saum; Albrecht Brandenburg; Christian Hoffmann (2007). "Noninvasive time-dependent cytometry monitoring by digital holography". Journal of Biomedical Optics. 12 (6): 064002. Bibcode:2007JBO....12f4002K. doi:10.1117/1.2804926. PMID 18163818. S2CID 40335328.
  4. ^ Myung K. Kim (2010). "Principles and techniques of digital holographic microscopy". SPIE Reviews. 1: 018005. Bibcode:2010SPIER...1a8005K. doi:10.1117/6.0000006.
  5. ^ Zangle T, Teitell M (2014). "Live-cell mass profiling: an emerging approach in quantitative biophysics". Nature Methods. 11 (12): 1221–1228. doi:10.1038/nmeth.3175. PMC 4319180. PMID 25423019.
  6. ^ Chen, Claire Lifan; Mahjoubfar, Ata; Tai, Li-Chia; Blaby, Ian K.; Huang, Allen; Niazi, Kayvan Reza; Jalali, Bahram (2016). "Deep Learning in Label-free Cell Classification". Scientific Reports. 6: 21471. Bibcode:2016NatSR...621471C. doi:10.1038/srep21471. PMC 4791545. PMID 26975219.published under CC BY 4.0 licensing
[edit]
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