Harmonic Generation Microscopy 2.0: New Tricks Empowering Intravital Imaging for Neuroscience

Lim, Henry C.

doi:10.3389/fmolb.2019.00099

Cited by 13 publications

(9 citation statements)

References 158 publications

(194 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Two-photon excited fluorescence microscopy is increasingly common in the vision research ( 35 , 72–78 ). In contrast, SHG microscopy, which has been widely used for structural imaging of collagen and myosin, is extended to the central nervous system only recently ( 79 ). Retinal SHG imaging can be performed using the same setup as two-photon fluorescence microscopy with minimum alterations, e.g., the forward signal detection.…”

Section: Discussionmentioning

confidence: 99%

Axonal architecture of the mouse inner retina revealed by second harmonic generation

et al. 2022

Self Cite

View full text Add to dashboard Cite

We describe a novel method for visualizing the network of axons in the unlabeled fresh wholemount retina. The intrinsic radiation of second harmonic generation (SHG) was utilized to visualize single axons of all major retinal neurons, i.e. photoreceptors, horizontal cells, bipolar cells, amacrine cells, and the retinal ganglion cells. The cell types of SHG + axons were determined using transgenic GFP/YFP mice. New findings were obtained with retinal SHG imaging: Müller cells do not maintain uniformly polarized microtubules in the processes; SHG + axons of bipolar cells terminate in the inner plexiform layer (IPL) in a subtype-specific manner; a subset of amacrine cells, presumably the axon-bearing types, emits SHG; and the axon-like neurites of amacrine cells provide a cytoskeletal scaffolding for the IPL stratification. To demonstrate the utility, retinal SHG imaging was applied for testing whether the inner retina is preserved in glaucoma, using DBA/2 mice as a model of glaucoma and DBA/2-Gpnmb + as the non-glaucomatous control. It was found that the morphology of the inner retina was largely intact in glaucoma and the pre-synaptic compartments to the retinal ganglion cells were uncompromised. It proves retinal SHG imaging as a promising technology for studying the physiological and diseased retinas in 3D.

show abstract

Section: Discussionmentioning

confidence: 99%

Axonal architecture of the mouse inner retina revealed by second harmonic generation

et al. 2022

Self Cite

View full text Add to dashboard Cite

show abstract

“…Thus, the result is a new photon with exactly double the energy of any of the two initial photons. Using this process as a contrast mechanism, SHG microscopy has become established as a powerful biomedical investigation technique, enabling to date a wide variety of biological and biophysical imaging applications 2 . Tubulin 3 , myosin 4 and collagen 5 , 6 are known to generate SHG signals, with the latter being one of the most frequently analysed tissue constituents by SHG microscopy.…”

Section: Background and Summarymentioning

confidence: 99%

PSHG-TISS: A collection of polarization-resolved second harmonic generation microscopy images of fixed tissues

et al. 2022

View full text Add to dashboard Cite

Second harmonic generation (SHG) microscopy is acknowledged as an established imaging technique capable to provide information on the collagen architecture in tissues that is highly valuable for the diagnostics of various pathologies. The polarization-resolved extension of SHG (PSHG) microscopy, together with associated image processing methods, retrieves extensive image sets under different input polarization settings, which are not fully exploited in clinical settings. To facilitate this, we introduce PSHG-TISS, a collection of PSHG images, accompanied by additional computationally generated images which can be used to complement the subjective qualitative analysis of SHG images. These latter have been calculated using the single-axis molecule model for collagen and provide 2D representations of different specific PSHG parameters known to account for the collagen structure and distribution. PSHG-TISS can aid refining existing PSHG image analysis methods, while also supporting the development of novel image processing and analysis methods capable to extract meaningful quantitative data from the raw PSHG image sets. PSHG-TISS can facilitate the breadth and widespread of PSHG applications in tissue analysis and diagnostics.

show abstract

“…When performing THG image analysis, care needs to be taken in the interpretation of the signal generation as THG is a coherent process. For further reading on application of THG and practicalities of SHG, Lim et al published a review on imaging intravital tissues using SHG and THG [105], and Chen et al published a comprehensive protocol for practical and quantitative SHG measurements [90].…”

Section: Understanding Intensities and Polarisation In Shgmentioning

confidence: 99%

Imaging approaches for monitoring three‐dimensional cell and tissue culture systems

Hilzenrat

Gill

McArthur

2022

Journal of Biophotonics

View full text Add to dashboard Cite

The past decade has seen an increasing demand for more complex, reproducible and physiologically relevant tissue cultures that can mimic the structural and biological features of living tissues. Monitoring the viability, development and responses of such tissues in real‐time are challenging due to the complexities of cell culture physical characteristics and the environments in which these cultures need to be maintained in. Significant developments in optics, such as optical manipulation, improved detection and data analysis, have made optical imaging a preferred choice for many three‐dimensional (3D) cell culture monitoring applications. The aim of this review is to discuss the challenges associated with imaging and monitoring 3D tissues and cell culture, and highlight topical label‐free imaging tools that enable bioengineers and biophysicists to non‐invasively characterise engineered living tissues.

show abstract

Harmonic Generation Microscopy 2.0: New Tricks Empowering Intravital Imaging for Neuroscience

Cited by 13 publications

References 158 publications

Axonal architecture of the mouse inner retina revealed by second harmonic generation

Axonal architecture of the mouse inner retina revealed by second harmonic generation

PSHG-TISS: A collection of polarization-resolved second harmonic generation microscopy images of fixed tissues

Imaging approaches for monitoring three‐dimensional cell and tissue culture systems

Contact Info

Product

Resources

About