Intravital microscopy: Difference between revisions
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==Basic Concept== |
==Basic Concept== |
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Intravital microscopy involves imaging cells of a live animal through an imaging window that is implanted into the animal tissue during a special surgery. The main advantage of intravital microscopy is that it allows imaging living cells while they are in the true environment of a complex multicellular organism. Thus, intravital microscopy allows researchers to study the behavior of cells in their natural environment or in vivo rather than in a cell culture. Another advantage of intravital microscopy is that the experiment can be set up in a way to allow observing changes in a living tissue of an organism over a period of time that is useful for many areas of research including cancer and stem cell research. |
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High quality of modern microscopes and imaging software also permits subcellular imaging in live animals that in turn allows studying cell biology at molecular level in vivo. Advancements in fluorescent protein technology and genetic tools that enable controlled expression of a given gene at a specific time in a tissue of interest also played important role in intravital microscopy development. |
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The possibility of generating appropriate transgenic mice is crucial for an intravital microscopy studies. For example, in order to study the behavior of microglial cells in Alzheimer’s disease researchers will need to crossbreed a transgenic mouse that is a mouse model of Alzheimer’s disease with another transgenic mouse that is a mouse model for visualization of microglial cells. Cells need to produce a fluorescent protein to be visualized and this can be achieved by introducing a transgene.<ref>{{cite journal|last1=Krabbe|first1=Grietje|last2=Halle|first2=Annett|last3=Matyash|first3=Vitali|last4=Rinnenthal|first4=Jan L.|last5=Eom|first5=Gina D.|last6=Bernhardt|first6=Ulrike|last7=Miller|first7=Kelly R.|last8=Prokop|first8=Stefan|last9=Kettenmann|first9=Helmut|last10=Heppner|first10=Frank L.|last11=Priller|first11=Josef|title=Functional Impairment of Microglia Coincides with Beta-Amyloid Deposition in Mice with Alzheimer-Like Pathology|journal=PLoS ONE|date=8 April 2013|volume=8|issue=4|pages=e60921|doi=10.1371/journal.pone.0060921}}</ref> |
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==Imaging== |
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If there is a need to capture close interactions between cells multiphoton microscopy is preferable as it provides greater depth of image than single-photon confocal microscopy. <ref name="Time-lapseIVM" /> |
If there is a need to capture close interactions between cells multiphoton microscopy is preferable as it provides greater depth of image than single-photon confocal microscopy. <ref name="Time-lapseIVM" /> |
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Multiphoton microscopy also allows visualization of cells located underneath bone tissues such as cells of the [[bone marrow]].<ref name="T-ALL">{{cite journal|last1=Hawkins|first1=Edwin D.|last2=Duarte|first2=Delfim|last3=Akinduro|first3=Olufolake|last4=Khorshed|first4=Reema A.|last5=Passaro|first5=Diana|last6=Nowicka|first6=Malgorzata|last7=Straszkowski|first7=Lenny|last8=Scott|first8=Mark K.|last9=Rothery|first9=Steve|last10=Ruivo|first10=Nicola|last11=Foster|first11=Katie|last12=Waibel|first12=Michaela|last13=Johnstone|first13=Ricky W.|last14=Harrison|first14=Simon J.|last15=Westerman|first15=David A.|last16=Quach|first16=Hang|last17=Gribben|first17=John|last18=Robinson|first18=Mark D.|last19=Purton|first19=Louise E.|last20=Bonnet|first20=Dominique|last21=Lo Celso|first21=Cristina|title=T-cell acute leukaemia exhibits dynamic interactions with bone marrow microenvironments|journal=Nature|date=17 October 2016|volume=538|issue=7626|pages=518–522|doi=10.1038/nature19801}}</ref> |
Multiphoton microscopy also allows visualization of cells located underneath bone tissues such as cells of the [[bone marrow]].<ref name="T-ALL">{{cite journal|last1=Hawkins|first1=Edwin D.|last2=Duarte|first2=Delfim|last3=Akinduro|first3=Olufolake|last4=Khorshed|first4=Reema A.|last5=Passaro|first5=Diana|last6=Nowicka|first6=Malgorzata|last7=Straszkowski|first7=Lenny|last8=Scott|first8=Mark K.|last9=Rothery|first9=Steve|last10=Ruivo|first10=Nicola|last11=Foster|first11=Katie|last12=Waibel|first12=Michaela|last13=Johnstone|first13=Ricky W.|last14=Harrison|first14=Simon J.|last15=Westerman|first15=David A.|last16=Quach|first16=Hang|last17=Gribben|first17=John|last18=Robinson|first18=Mark D.|last19=Purton|first19=Louise E.|last20=Bonnet|first20=Dominique|last21=Lo Celso|first21=Cristina|title=T-cell acute leukaemia exhibits dynamic interactions with bone marrow microenvironments|journal=Nature|date=17 October 2016|volume=538|issue=7626|pages=518–522|doi=10.1038/nature19801}}</ref> |
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Revision as of 14:50, 22 March 2018
 | This article needs additional citations for verification. (June 2013) |
Intravital microscopy is a form of microscopy that allows observing biological processes in live animals (in vivo) at a high resolution that makes distinguishing between individual cells of a tissue possible. [1] [2]
Before an animal can be used for intravital microscopy imaging it has to undergo a surgery involving implantation of an imaging window. For example, if researchers want to visualize liver cells of a live mouse they will implant an imaging window into mouse’s abdomen.[3]
Mice are the most common choice of animals for intravital microscopy but in special cases other rodents such as rats might be more suitable. Animals are always anesthetized throughout surgeries and imaging sessions. [2]
Intravital microscopy is used in several areas of research including neurology, immunology, stem cell and others. This technique is particularly useful to assess a progression of a disease or an effect of a drug. [2]
Basic Concept
Intravital microscopy involves imaging cells of a live animal through an imaging window that is implanted into the animal tissue during a special surgery. The main advantage of intravital microscopy is that it allows imaging living cells while they are in the true environment of a complex multicellular organism. Thus, intravital microscopy allows researchers to study the behavior of cells in their natural environment or in vivo rather than in a cell culture. Another advantage of intravital microscopy is that the experiment can be set up in a way to allow observing changes in a living tissue of an organism over a period of time that is useful for many areas of research including cancer and stem cell research. High quality of modern microscopes and imaging software also permits subcellular imaging in live animals that in turn allows studying cell biology at molecular level in vivo. Advancements in fluorescent protein technology and genetic tools that enable controlled expression of a given gene at a specific time in a tissue of interest also played important role in intravital microscopy development.
The possibility of generating appropriate transgenic mice is crucial for an intravital microscopy studies. For example, in order to study the behavior of microglial cells in Alzheimer’s disease researchers will need to crossbreed a transgenic mouse that is a mouse model of Alzheimer’s disease with another transgenic mouse that is a mouse model for visualization of microglial cells. Cells need to produce a fluorescent protein to be visualized and this can be achieved by introducing a transgene.[4]
Imaging
If there is a need to capture close interactions between cells multiphoton microscopy is preferable as it provides greater depth of image than single-photon confocal microscopy. [5] Multiphoton microscopy also allows visualization of cells located underneath bone tissues such as cells of the bone marrow.[6]
Limitations of Intravital Microscopy
One of the main advantages of intravital microscopy is the opportunity to observe how cells interact with their microenvironment. However, visualization of all types of cells in the microenvironment is limited by the number of distinguishable fluorescent labels available.[5] It is also widely accepted that some tissues such as brain can be visualized easier than others such as skeletal muscle. [2] In addition, generating transgenic mice with a phenotype of interest and fluorescent proteins in appropriate cell types is often challenging and time consuming.[5]
References
- ^ Gavins, Felicity N.E.; Chatterjee, Bristi E. (2004). "Intravital microscopy for the study of mouse microcirculation in anti-inflammatory drug research: Focus on the mesentery and cremaster preparations". Journal of Pharmacological and Toxicological Methods. 49: 1–14. doi:10.1016/S1056-8719(03)00057-1.
- ^ a b c d Masedunskas, Andrius; Milberg, Oleg; Porat-Shliom, Natalie; Sramkova, Monika; Wigand, Tim; Amornphimoltham, Panomwat; Weigert, Roberto. "Intravital microscopy A practical guide on imaging intracellular structures in live animals". Bioarchitecture. 2 (5): 143–157. doi:10.4161/bioa.21758. PMC 3696059. PMID 22992750.
- ^ Ritsma, Laila; Steller, Ernst J. A.; Beerling, Evelyne; Loomans, Cindy J. M.; Zomer, Anoek; Gerlach, Carmen; Vrisekoop, Nienke; Seinstra, Daniëlle; Gurp, Leon van; Schäfer, Ronny; Raats, Daniëlle A.; Graaff, Anko de; Schumacher, Ton N.; Koning, Eelco J. P. de; Rinkes, Inne H. Borel; Kranenburg, Onno; Rheenen, Jacco van (31 October 2012). "Intravital Microscopy Through an Abdominal Imaging Window Reveals a Pre-Micrometastasis Stage During Liver Metastasis". Science Translational Medicine. 4 (158): 158ra145–158ra145. doi:10.1126/scitranslmed.3004394. ISSN 1946-6234.
- ^ Krabbe, Grietje; Halle, Annett; Matyash, Vitali; Rinnenthal, Jan L.; Eom, Gina D.; Bernhardt, Ulrike; Miller, Kelly R.; Prokop, Stefan; Kettenmann, Helmut; Heppner, Frank L.; Priller, Josef (8 April 2013). "Functional Impairment of Microglia Coincides with Beta-Amyloid Deposition in Mice with Alzheimer-Like Pathology". PLoS ONE. 8 (4): e60921. doi:10.1371/journal.pone.0060921.
{{cite journal}}: CS1 maint: unflagged free DOI (link) - ^ a b c Harney, Allison S.; Wang, Yarong; Condeelis, John S.; Entenberg, David (12 June 2016). "Extended Time-lapse Intravital Imaging of Real-time Multicellular Dynamics in the Tumor Microenvironment". Journal of Visualized Experiments (112). doi:10.3791/54042.
- ^ Hawkins, Edwin D.; Duarte, Delfim; Akinduro, Olufolake; Khorshed, Reema A.; Passaro, Diana; Nowicka, Malgorzata; Straszkowski, Lenny; Scott, Mark K.; Rothery, Steve; Ruivo, Nicola; Foster, Katie; Waibel, Michaela; Johnstone, Ricky W.; Harrison, Simon J.; Westerman, David A.; Quach, Hang; Gribben, John; Robinson, Mark D.; Purton, Louise E.; Bonnet, Dominique; Lo Celso, Cristina (17 October 2016). "T-cell acute leukaemia exhibits dynamic interactions with bone marrow microenvironments". Nature. 538 (7626): 518–522. doi:10.1038/nature19801.
External links
Media related to Intravital microscopy at Wikimedia Commons
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