Night in Tunisia, Part 1 Gmaj.7 Em 87-9 Em Em Em Em 87-5 ...
Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob...
Transcript of Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob...
![Page 1: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/1.jpg)
18-12-2017
Ben N. G. Giepmans Dept. Cell Biology, UMCG Microscopy & Imaging Center (UMIC) UMC Groningen, The Netherlands [email protected] www.cellbiology.nl
Nanotomy &
High-Resolution Color-EM
![Page 2: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/2.jpg)
Nanotomy &
High-Resolution Color-EM
Jacob Hoogenboom
Marijke Scotuzzi
Kees Hagen
Dasha Wensveen
Ben Giepmans
Jeroen Kuipers
Pascal de Boer
![Page 3: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/3.jpg)
Probes & techniques for microscopy
- Identify and quantify -
How do molecules, organelles & cells regulate (patho)physiology Need probes: Resolution, dynamics, identification, quantification Large scale EM / nano-anatomy (“nanotomy”): More identification/quantification at high resolution Big data sharing New probes & techniques allow nm-scale “color-EM”: Proof-of-concept and limitations
![Page 5: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/5.jpg)
Large scale EM (Nanotomy)
5 rbv3.com/blog/wp-content/uploads/2014/03/Google.jpg nanotomy.org
Nano-anatomy: anatomy of tissues, cells, organelles at the macromolecular level, “Google-tissue” Major benefits of Nanotomy: - Overview and high resolution - Rare events may be imaged - Unbiased examination of tissue, cells, organelles and molecules - Open access raw data sharing - Routine technique - Grey scale images www.nanotomy.org
![Page 6: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/6.jpg)
175 µm 64 µm 16 µm 4 µm 1 µm
From Islet to Insulin granule
![Page 7: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/7.jpg)
7 https://pdb101.rcsb.org/motm/14
Insulin
![Page 8: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/8.jpg)
8 http://chemistry.tutorvista.com/biochemistry/cysteine.html
Cysteine
![Page 9: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/9.jpg)
9 Shutterstock
Sulphur
![Page 10: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/10.jpg)
Energy dispersive X-rays (EDX)
10 Thermo Scientific
Incident electrons
Image by: Thermo Scientific
![Page 11: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/11.jpg)
11 Bushberg et al. (2011)
Energy dispersive X-rays (EDX)
![Page 12: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/12.jpg)
Energy dispersive X-rays (EDX)
12 pinterest.com
![Page 13: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/13.jpg)
X-ray detection
13
X-ray detector
Hoogenboom et al., TU-Delft
![Page 14: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/14.jpg)
![Page 15: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/15.jpg)
15 www.bpc.edu
Periodic table of elements
![Page 16: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/16.jpg)
X-ray spectrum of gold versus cadmium
16 www.xrfresearch.com
![Page 17: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/17.jpg)
Detect Au in immunogold and Cd in QDs?
Scotuzzi et al. Scientific Reports 2017
![Page 18: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/18.jpg)
EDX reveals Cd in QDs labeling insulin
18 Scotuzzi et al. Scientific Reports 2017
![Page 19: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/19.jpg)
Au /G4 at P-rich heterochromatin
19 Scotuzzi et al. Scientific Reports 2017
![Page 20: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/20.jpg)
Multi-colour nm-scale EM (G4 and insulin)
Scotuzzi et al. Scientific Reports 2017
![Page 21: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/21.jpg)
Multi-colour nm-scale EM (G4 and insulin)
Scotuzzi et al. Scientific Reports 2017
![Page 22: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/22.jpg)
Energy-Dispersive X-ray Spectra (EDX / EDS)
22 Scotuzzi et al. Scientific Reports 2017
![Page 23: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/23.jpg)
Elemental maps
23
Au Ca Cd Cl Fe
K Mg N Na Os
P S Se U Zn
Scotuzzi et al. Scientific Reports 2017
![Page 24: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/24.jpg)
Contrast /morphology: Distinct granules?
24 Scotuzzi et al. Scientific Reports 2017
![Page 25: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/25.jpg)
Distinct granules based on elements!
25 Scotuzzi et al. Scientific Reports 2017
![Page 26: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/26.jpg)
Distinct granules based on elements!
26 Scotuzzi et al. Scientific Reports 2017
![Page 27: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/27.jpg)
18-12-2017
Scotuzzi et al. Scientific Reports 2017
![Page 28: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/28.jpg)
Nanotomy, EDX & ColorEM
“Superdepuper” resolution colorEM with EDX
Endogenous elements (S; P; distinct vesicles)
Paint organelles (Os, W)
Identify labels (Cd, Au)
Revolution in computer sciences allow data
acquisition and data sharing
Large-scale EM
www.nanotomy.org
Scie
nce
(20
06
)
![Page 29: Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob Hoogenboom Marijke Scotuzzi Kees Hagen Dasha Wensveen Ben Giepmans Jeroen Kuipers Pascal de](https://reader033.fdocuments.us/reader033/viewer/2022051720/5a78cff37f8b9aa17b8cef06/html5/thumbnails/29.jpg)
Acknowledgements
University Medical Center Groningen
Jeroen Kuipers
Pascal de Boer
Anouk Wolters
Marit de Beer
Nicole Pirozzi
Klaas Sjollema
Delft University of Technology
Marijke Scotuzzi
Dasha Wensveen
Lennard Voortman
Kees Hagen
Jacob Hoogenboom