Learn & Share: Multiphoton Microscopy
With multiphoton microscopy we can image several hundreds of micrometers deep into biological samples as scattering of red-shifted light in tissues is highly reduced compared to visible light. For multiphoton excitation, pulsed infrared lasers with wavelengths of up to 1300 nm in case of the OPO (optical-parametric oscillator, What is an OPO?) are used.
Deep tissue imaging of thick specimens or in whole animals allows to observe cellular processes in their natural context. Especially intravital imaging with multiphoton excitation plays a growing role in many biomedical research areas.
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Image Gallery: Tunable Multicolor Deep In Vivo Imaging
Today’s life science research focusses on complex biological processes, such as the causes of cancer and other human diseases. A deep look into tissues and living specimens is vital to understanding…Read article -
Direct visualization of the arterial wall water permeability barrier using CARS microscopy
The artery wall is equipped with a water permeation barrier that allows blood to flow at high pressure without significant water leak. The precise location of this barrier is unknown despite its…Read article -
High-Resolution 3D Imaging of Whole Organ after Clearing
Zebrafish testis has become a powerful model for reproductive biology of teleostean fishes and other vertebrates and encompasses multiple applications in applied and basic research. Many studies have…Read article -
Multiphoton Microscopy – a Satisfied Wish List
The colorful picture shows colon tumor cells, fluorescently labelled and lineage traced from a multicolor tracer. The gray color codes for the second harmonic generation (SHG) signal from Collagen 1.…Read article -
Mission Impossible Accomplished: Tunable Colors for Non-descanning Detection
Leica Microsystems’ 4Tune detector, the key component of the SP8 DIVE Deep In Vivo Explorer, provides spectrally tunable image recording with non-descanning detection. An innovative solution for…Read article -
Laser Beam Shaping for Multicolor Multiphoton Microscopy
Multiphoton Microscopy is one of the current hot topics in life science research. The new Leica TCS SP8 DIVE from Leica Microsystems presents a series of beneficial new innovations, including a freely…Read article -
Five Questions Asked: Prof. Dr. Jacco van Rheenen speaks about the most important considerations when imaging deep into mouse tissue
When operating a confocal microscope, or when discussing features and parameters of such a device, we inescapably mention the pinhole and its diameter. This short introductory document is meant to…Read article -
Chronic Stress in Mice Remodels Lymph Vasculature to Promote Tumour Cell Dissemination
Chronic stress induces signalling from the sympathetic nervous system (SNS) and drives cancer progression, although the pathways of tumour cell dissemination are unclear. Here we show that chronic…Read article -
Clarifying Tissue Clearing
Biological specimens are intrinsically three dimensional; however because of the obscuring effects of light scatter, imaging deep into a tissue volume is problematic. Although efforts to eliminate the…Read article -
Multiphoton Microscopy Publication List
Multiphoton Microscopy is an advanced technique for imaging thick samples. Applications range from the visualization of the complex architecture of the whole brain to the study of tumor development…Read article -
BABB Clearing and Imaging for High Resolution Confocal Microscopy: Counting and Sizing Kidney Cells in the 21st Century
Multipohoton microscopy experiment using Leica TCS SP8 MP and Leica 20x/0.95 NA BABB immersion objective. Understanding kidney microanatomy is key to detecting and identifying early events in kidney…Read article -
Clearing of Fixed Tissue: A Review from a Microscopist’s Perspective
Chemical clearing of fixed tissues is becoming a key instrument for the three-dimensional reconstruction of macroscopic tissue portions, including entire organs. Indeed, the growing interest in this…Read article -
4Pi-RESOLFT Nanoscopy
Here we apply the 4Pi scheme to RESOLFT nanoscopy using two-photon absorption for the on-switching of fluorescent proteins. We show that in this combination, the lobes are so low that low-light level,…Read article -
From Light to Mind: Sensors and Measuring Techniques in Confocal Microscopy
This article outlines the most important sensors used in confocal microscopy. By confocal microscopy, we mean "True Confocal Scanning", i.e. the technique that illuminates and measures one single…Read article -
Correlating Intravital Multi-Photon Microscopy to 3D Electron Microscopy of Invading Tumor Cells Using Anatomical Reference Points
Cancer research unsing multiphoton microscopy and 3D electron microscopy. Correlative microscopy combines the advantages of both light and electron microscopy to enable imaging of rare and transient…Read article -
Third Harmonic Generation Microscopy – Label-Free 3D-Tissue Imaging and Blood Flow Characterization
THG microscopy as special variants of multiphoton microscopy. Third Harmonic Generation (THG) microscopy is a non-fluorescent multi-photon technique that combines the advantages of label-free…Read article -
Interview with Dr. Gertrude Bunt and Prof. Fred S. Wouters on the FOM 2015
Only a few days to go before the start of Focus on Microscopy 2015 in Göttingen, Germany. This year’s FOM is being organized by Dr. Gertrude Bunt and Prof. Dr. Fred S. Wouters from the University…Read article -
How to Choose the Right Confocal Microscope for Your Lab?
Confocal microscopy has come a very long way since its invention more than a half-century ago. Today, with novel technology driven by leading imaging companies, it has become the standard for…Read article -
Deeper Insights in Transparent Animals
CLARITY clearing derivatives for multiphoton microscopy. Transparent organisms help us to identify spatial arrangements and connections of cells and tissues, especially neuronal circuits can easily…Read article -
Clearing Procedures for Deep Tissue Imaging
Multi-channel multiphoton microscopy with dedicated optics for CLARITY. Why clearing? Curiosity is human nature. And nothing attracts as much curiosity as the inside of living organisms. While in…Read article -
Single-Cell Phenotyping within Transparent Intact Tissue through Whole-Body Clearing
Understanding the structure-function relationships at cellular, circuit, and organ-wide scale requires 3D anatomical and phenotypical maps, currently unavailable for many organs across species. At the…Read article -
The Environment Makes the Stem Cell
A recent publication in Nature shows that all stem cells divide and compete for niche space by passively "kicking out" others so that eventually one stem cell takes over the whole niche. Jacco van…Read article -
Smart Control for Resonant Galvo Scanners
High time-resolution confocal microscopy (HTRCLSM) requires fast scanning devices. Whereas non-resonant galvo scanners allow full position control, but only at slow speed, resonant scanners allow…Read article -
Tracking Glomerular Fate Over Long Time Distances
Multi-channel multiphoton microscopy with dedicated optics for CLARITY. The glomerular filtration barrier (GFB) is a complex spatial structure within the kidney glomerulis where ultrafiltration…Read article -
Map the Brain with CLARITY
Imaging whole brains with CLARITY and multiphoton microscopy. Image a whole brain without sectioning? Investigate neuronal circuits without reconstruction? Perform molecular phenotyping without…Read article -
Single-Wavelength Two-Photon Excitation-stimulated Emission Depletion (SW2PE-STED) Superresolution Imaging
Two-photon microscopy, multiphoton microcopy and super-resolution imaging. We developed a new class of two-photon excitation–stimulated emission depletion (2PE-STED) optical microscope. In this…Read article -
Direct In Vivo Evidence for Tumor Propagation by Glioblastoma Cancer Stem Cells
Cancer research using multiphoton microscopy. High-grade gliomas (World Health Organization grade III anaplastic astrocytoma and grade IV glioblastoma multiforme), the most prevalent primary…Read article -
Detectors for Sensitive Detection: HyD
This article discusses detectors (more precisely: sensors), that are employed in single point, i.e. true confocal scanning microscopes. The sensors in such systems are usually photomultiplier tubes.…Read article -
Developments in Multiphoton Excitation Microscopy
Basics, history, and applications of multiphoton microscopy. Honouring Goeppert-Mayer’s prediction of simultaneous two-photon absorption by an atom or molecule reported in the 1930s in her PhD…Read article -
Principles of Multiphoton Microscopy for Deep Tissue Imaging
Basics of multiphoton microscopy. This interactive tutorial explains the principles of multiphoton microscopy for deep tissue imaging. Multiphoton microscopy uses excitation wavelengths in the…Read article
Interactive Tutorials
Useful Links
Communities and Web Sources
www.researchgate.net/Social network for scientists
www.lsoft.com/scripts/wl.exe?SL1=CONFOCALMICROSCOPY&H=LISTS.UMN.EDUConfocal Microscopy Mailing List, University of Minnesota
www.ibiology.org/Teaching tools, video lectures on biology and microscopy
bitesizebio.comOnline magazine and community for molecular and cell biology researchers
www.somersault1824.comResource for high-end scientific illustrations, images and animations
Search Engines and Data Bases
www.cellimagelibrary.orgPublic resource database of images, videos, and animations of cells
harvester.fzk.de/harvesterBioinformatic meta search engine for genes and proteins
www.gopubmed.comSearch interface for pubmed
en.wikipedia.org/wiki/List_of_academic_databases_and_search_enginesList of academic databases and search engines
scholar.google.comBeta of Google's search engine for scientific article abstracts
Journals
www.doaj.org/Directory of open access journals
emboj.embopress.org/The EMBO Journal
www.lifescied.orgCBE-Life Sciences Education – an ASCB online journal
www.sciencemag.org/Science
www.nature.com/Nature
www.cell.com/Biweekly publication of exceptional research articles
jcs.biologists.org/Journal of Cell Science
dev.biologists.org/Development
jeb.biologists.org/The Journal of Experimental Biology
dmm.biologists.org/DMM Disease Models & Mechanisms
www.biotechniques.com/International Journal of Life Science Methods
www.opticsinfobase.org/Collection of Journals and Proceedings in Optics and Photonics
spie.org/x576.xmlSPIE - peer-reviewed journals on applied research in optics and photonics
onlinelibrary.wiley.com/journal/10.1002/(ISSN)1864-0648Journal of Biophotonics
www.plosone.org/home.actionInternational, peer-reviewed, open-access, online publication
rspb.royalsocietypublishing.org/Proceedings B - the Royal Society's biological research journal
www.microscopy-analysis.com/International Journal for microscopists
Organizations
www.microscopy.org/Microscopy Society of America
www.eurmicsoc.org/European Microscopy Society
www.rms.org.uk/Royal Microscopical Society
www.ascb.org/ASCB American Society of Cell Biology
www.biologists.com/cob_activities.htmlthe company of biologists
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