English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Visualization of bacterial protein complexes labeled with fluorescent proteins and nanobody binders for STED microscopy

Cramer, K., Bolender, A.-L., Stockmar, I., Jungmann, R., Kasper, R., & Shin, J. Y. (2019). Visualization of bacterial protein complexes labeled with fluorescent proteins and nanobody binders for STED microscopy. International Journal of Molecular Sciences, 20(14): 3376. doi:10.3390/ijms20143376.

Item is

Basic

show hide
Genre: Journal Article

Files

show Files
hide Files
:
ijms-20-03376.pdf (Publisher version), 2MB
Name:
ijms-20-03376.pdf
Description:
-
OA-Status:
Visibility:
Public
MIME-Type / Checksum:
application/pdf / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited
:
ijms-20-03376-s001.pdf (Supplementary material), 449KB
Name:
ijms-20-03376-s001.pdf
Description:
-
OA-Status:
Visibility:
Public
MIME-Type / Checksum:
application/pdf / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-

Locators

show

Creators

show
hide
 Creators:
Cramer, Kimberly1, Author           
Bolender, Anna-Lena2, Author           
Stockmar, Iris1, Author           
Jungmann, Ralf1, Author           
Kasper, Robert3, Author           
Shin, Jae Yen1, Author           
Affiliations:
1Jungmann, Ralf / Molecular Imaging and Bionanotechnology, Max Planck Institute of Biochemistry, Max Planck Society, ou_2149679              
2Department: Molecules-Signaling-Development / Klein, MPI of Neurobiology, Max Planck Society, ou_1113546              
3MPI of Neurobiology, Max Planck Society, ou_1110547              

Content

show
hide
Free keywords: ACTIVATED LOCALIZATION MICROSCOPY; SUPERRESOLUTION MICROSCOPY; RESOLUTION LIMIT; CELL-DIVISION; SINGLE; SUBTILIS; FTSZBiochemistry & Molecular Biology; Chemistry; STED; bacteria; super-resolution microscopy; fluorescent proteins; nanobody; cell division;
 Abstract: In situ visualization of molecular assemblies near their macromolecular scale is a powerful tool to investigate fundamental cellular processes. Super-resolution light microscopies (SRM) overcome the diffraction limit and allow researchers to investigate molecular arrangements at the nanoscale. However, in bacterial cells, visualization of these assemblies can be challenging because of their small size and the presence of the cell wall. Thus, although conceptually promising, successful application of SRM techniques requires careful optimization in labeling biochemistry, fluorescent dye choice, bacterial biology and microscopy to gain biological insights. Here, we apply Stimulated Emission Depletion (STED) microscopy to visualize cell division proteins in bacterial cells, specifically E. coli and B. subtilis. We applied nanobodies that specifically recognize fluorescent proteins, such as GFP, mCherry2 and PAmCherry, fused to targets for STED imaging and evaluated the effect of various organic fluorescent dyes on the performance of STED in bacterial cells. We expect this research to guide scientists for in situ macromolecular visualization using STED in bacterial systems.

Details

show
hide
Language(s): eng - English
 Dates: 2019-07-10
 Publication Status: Published in print
 Pages: 11
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Identifiers: ISI: 000480449300001
DOI: 10.3390/ijms20143376
 Degree: -

Event

show

Legal Case

show

Project information

show hide
Project name : (DFG JU 2957/1-1 to R.J.)
Grant ID : -
Funding program : Emmy Noether Program
Funding organization : German Research Foundation
Project name : MolMap (grant agreement number 680241 to R.J.)
Grant ID : 680241
Funding program : ERC Starting Grant
Funding organization : European Research Council

Source 1

show
hide
Title: International Journal of Molecular Sciences
  Abbreviation : Int. J. Mol. Sci.
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: Basel, Switzerland : MDPI AG
Pages: - Volume / Issue: 20 (14) Sequence Number: 3376 Start / End Page: - Identifier: ISSN: 1422-0067
CoNE: https://pure.mpg.de/cone/journals/resource/1422-0067