Aqueous mounting media increasing tissue translucence improve image quality in Structured Illumination Microscopy of thick biological specimen

Scientific Reports
Aleksander SzczurekSabine Mai

Abstract

Structured Illumination Microscopy (SIM) is a super-resolution microscopy method that has significantly advanced studies of cellular structures. It relies on projection of illumination patterns onto a fluorescently labelled biological sample. The information derived from the sample is then shifted to a detectable band, and in the process of image calculation in Fourier space the resolution is doubled. Refractive index homogeneity along the optical path is crucial to maintain a highly modulated illumination pattern necessary for high-quality SIM. This applies in particular to thick samples consisting of large cells and tissues. Surprisingly, sample mounting media for SIM have not undergone a significant evolution for almost a decade. Through identification and systematic evaluation of a number of non-hazardous, water-soluble chemical components of mounting media, we demonstrate an unprecedented improvement in SIM-image quality. Mounting solutions presented in this research are capable of reducing abundant light scattering which constitutes the limiting factor in 3D-SIM imaging of large Hodgkin's lymphoma and embryonic stem cells as well as 10 µm tissue sections. Moreover, we demonstrate usefulness of some of the media in single ...Continue Reading

References

Apr 1, 1968·Journal of Theoretical Biology·S Oki
Jul 9, 2004·Biophysical Journal·Janos Vörös
Nov 30, 2006·Microscopy Research and Technique·Thorsten StaudtStefan W Hell
Jun 26, 2008·Optics Express·Delphine DébarreTony Wilson
Nov 20, 2009·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Philbert S TsaiDavid Kleinfeld
Jul 23, 2011·Journal of Microscopy·A Ustione, D W Piston
Mar 16, 2012·Optics Express·Muthuvel ArigovindanDavid A Agard
Jun 30, 2012·Nature Methods·Johannes SchindelinAlbert Cardona
Sep 6, 2012·Cellular Microbiology·Carla Coltharp, Jie Xiao
Jun 14, 2013·Biomedical Optics Express·Nicolas OlivierSuliana Manley
Jul 31, 2013·The Journal of Cell Biology·Sébastien BrittonStephen P Jackson
Nov 26, 2014·Nature Methods·Vivien Marx
Mar 12, 2015·Frontiers in Neuroanatomy·Bing HouTianzi Jiang
Jul 4, 2015·Histochemistry and Cell Biology·Anne BurgertMarkus Sauer
Jul 18, 2015·Cell·Douglas S Richardson, Jeff W Lichtman
Nov 4, 2015·Scientific Reports·Graeme BallLothar Schermelleh
Nov 28, 2015·BMC Developmental Biology·Jens PopkenValeri Zakhartchenko
Nov 10, 2016·Optics Express·A MalavalliC M Aegerter
Apr 14, 2017·Nature Protocols·Felix KrausYolanda Markaki
Apr 14, 2017·Nature Protocols·Justin DemmerleLothar Schermelleh

❮ Previous
Next ❯

Citations

Apr 17, 2020·Scientific Reports·Aleksandra ArsićIvana Nikić-Spiegel
Jul 3, 2021·International Journal of Molecular Sciences·Paweł MatrybaJakub Gołąb
Aug 27, 2021·Current Protocols·Jessica Valli, Jeremy Sanderson

❮ Previous
Next ❯

Methods Mentioned

BETA
super-resolution microscopy
light scattering
fluorescence imaging
electron microscopy

Software Mentioned

ImageJ
SRRF ImageJ
ZEN
ThunderSTORM
FiJi
SIM
TDE97
NanoJ
SIMcheck

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.