High-resolution Imaging of Cleared and Expanded Kidney Tissue Samples

↓ Direkt till sidans innehåll
↓ Direkt till sidans sekundära innehåll (sidomenyn)

Sökning: WFRF:(Jess David Unnersjö) > High-resolution Ima...

Jess, David UnnersjöKTH,Biofysik,Cell Physics (författare)

High-resolution Imaging of Cleared and Expanded Kidney Tissue Samples

BokEngelska2019

Förlag, utgivningsår, omfång ...

KTH Royal Institute of Technology,2019
85 s.
electronicrdacarrier

Nummerbeteckningar

LIBRIS-ID:oai:DiVA.org:kth-249002
ISBN:9789178731633
https://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-249002URI

Kompletterande språkuppgifter

Språk:engelska
Sammanfattning på:engelska

Ingår i deldatabas

SwePubSwePub

Klassifikation

Ämneskategori:vet swepub-contenttype
Ämneskategori:dok swepub-publicationtype

Serie

TRITA-SCI-FOU ;2019:18

Anmärkningar

QC 20190411
The kidney is one of the most important and complex organs in the humanbody with the task of filtering hundreds of litres of blood daily. It is responsiblefor the salt and acid/base balance in the body, as well as secretinghormones important for red blood cell production and blood pressure regulation. Kidney disease is one of the fastest growing causes of death in the modern world, and this motivates extensive research for better understandingthe function of the kidney in both health and disease. Kidney failure or end stage renal disease (ESRD) is irreversible and requires treatment with dialysisor transplantation. Some of the most important cellular structures for blood filtration in the kidney are of very small dimensions (below 200 nanometers), and thus electron microscopy has previously been the only method with high enough resolution to study the morphology and topology of these minute structures. In three studies included in this thesis, we show that the finest elements of the kidney can now be resolved using different light microscopy techniques. In study 1, we show that by combining optical clearing with STED microscopy, protein localizations in the slit diaphragm of the kidney can be resolved, with widths around 75 nanometers. In study 3, a novel sample preparation method, expansion microscopy, is utilized to isotropically expand kidney tissue samples in space. Expansion improves the effective resolution by a factor of 5, making it possible to resolve podocyte foot processes and the slit diaphragmusing diffraction-limited confocal microscopy. We also show that by combining expansion microscopy and STED microscopy, the effective resolution can be improved even further (<20 nm). In our most recent work, study 5, we apply a simplified, moderate tissue swelling protocol which together with optimization of the confocal imaging provides sufficient resolution to resolve foot processes and parts of the filtration barrier. This new protocol is fast and technically simple, making it ideal for routine use, such as for future clinical pathology. In collaboration with kidney researchers, we have applied both STED microscopy and expansion microscopy to various disease models, showing that these tools can be used to both visualize and quantify pathologies occurring in different parts of the glomerular filtration barrier (GFB). In study 2, STED microscopy in combination with optical clearing is used to study the localization of Coro2b in secondary foot processes in both mouse and human tissue. In two ongoing studies with preliminary results presented in the thesis, we use STED microscopy and optical clearing to study the pathogenesis of focal segmental glomerulosclerosis (FSGS) by the use of genetic mouse models. Based on STED images, we extract different morphological parameters from foot processes and the glomerular filtration barrier (GFB) at different stages of the disease. In study 4, we apply a tissue expansion protocol to answer questions about the phenotype seen in podocytes where the mediator complex subunit 22 (Med22) is inactivated. By inactivating Med22 in a transgenic mouse line with cytosolic expression of tdTomato in podocytes, we saw strong indications that the vesicle-like structures seen in EM micrographs were indeed intracellular vesicles and not dilated sub-podocyte space. In summary, the work presented in this thesis has contributed to the development of a new toolbox for imaging renal ultra-structure using light microscopy, a field previously reserved for electron microscopy.

Ämnesord och genrebeteckningar

NATURVETENSKAP Biologi Biofysik hsv//swe
NATURAL SCIENCES Biological Sciences Biophysics hsv//eng
NATURVETENSKAP Biologi Cellbiologi hsv//swe
NATURAL SCIENCES Biological Sciences Cell Biology hsv//eng
MEDICIN OCH HÄLSOVETENSKAP Medicinska och farmaceutiska grundvetenskaper Cell- och molekylärbiologi hsv//swe
MEDICAL AND HEALTH SCIENCES Basic Medicine Cell and Molecular Biology hsv//eng
MEDICIN OCH HÄLSOVETENSKAP Klinisk medicin Urologi och njurmedicin hsv//swe
MEDICAL AND HEALTH SCIENCES Clinical Medicine Urology and Nephrology hsv//eng
TEKNIK OCH TEKNOLOGIER Medicinteknik Medicinsk laboratorie- och mätteknik hsv//swe
ENGINEERING AND TECHNOLOGY Medical Engineering Medical Laboratory and Measurements Technologies hsv//eng
Kidney
Renal Pathology
Microscopy
Super-resolution microscopy
Tissue expansion
Foot processes
Glomerular filtration
Podocytes
Biological Physics
Biologisk fysik

Biuppslag (personer, institutioner, konferenser, titlar ...)

Blom, Hans,1968-KTH,Biofysik(Swepub:kth)u1qhk3ta (preses)
Brismar, HjalmarKTH,Biofysik(Swepub:kth)u1232ew5 (preses)
Scott, Lena,Assistant ProfessorDept. of Women's and children's health, Karolinska Institutet (preses)
Endlich, KarlhansDept. of Anatomy and Cell Biology, University Medicine of Greifswald, Greifswald, Germany (opponent)
KTHBiofysik (creator_code:org_t)

Internetlänk

Hitta via bibliotek

High-resolution Imaging of Cleared and Expanded Kidney Tissue Samples (Sök publikationen i LIBRIS)

Till lärosätets databas

Hitta mer i SwePub

Av författaren/redakt...: Jess, David Unne ...; Blom, Hans, 1968 ...; Brismar, Hjalmar; Scott, Lena, Ass ...; Endlich, Karlhan ...

Om ämnet

NATURVETENSKAP: NATURVETENSKAP; och Biologi; och Biofysik

NATURVETENSKAP: NATURVETENSKAP; och Biologi; och Cellbiologi

MEDICIN OCH HÄLSOVETENSKAP: MEDICIN OCH HÄLS ...; och Medicinska och f ...; och Cell och molekyl ...

MEDICIN OCH HÄLSOVETENSKAP: MEDICIN OCH HÄLS ...; och Klinisk medicin; och Urologi och njur ...

TEKNIK OCH TEKNOLOGIER: TEKNIK OCH TEKNO ...; och Medicinteknik; och Medicinsk labora ...

Delar i serien: TRITA-SCI-FOU ;

Av lärosätet: Kungliga Tekniska Högskolan

Sök utanför SwePub

Sök vidare i:: Google; Google Book Search; Google Scholar

Kungliga biblioteket hanterar dina personuppgifter i enlighet med EU:s dataskyddsförordning (2018), GDPR. Läs mer om hur det funkar här.
Så här hanterar KB dina uppgifter vid användning av denna tjänst.

LIBRIS.kb.se