Probing endoplasmic reticulum dynamics using fluorescence imaging and photobleaching techniques.

Lindsey Costantini, Erik Snapp

Research output: Contribution to journalArticle

Abstract

This unit describes approaches and tools for studying the dynamics and organization of endoplasmic reticulum (ER) membranes and proteins in living cells using fluorescence microscopy. The ER plays a key role in secretory protein biogenesis, calcium regulation, and lipid synthesis. However, study of these processes has often been restricted to biochemical assays that average millions of lysed cells or imaging of static fixed cells. With new fluorescent protein (FP) reporter tools, sensitive commercial microscopes, and photobleaching techniques, investigators can interrogate the behaviors of ER proteins, membranes, and stress pathways in single live cells. Solutions are described for imaging challenges relevant to the ER, including the mobility of ER membranes, a range of ER structures, and the influence of post-translational modifications on FP reporters. Considerations for performing photobleaching assays for ER proteins are discussed. Finally, reporters and drugs for studying misfolded secretory protein stress and the unfolded protein response are described.

Original languageEnglish (US)
JournalCurrent protocols in cell biology / editorial board, Juan S. Bonifacino ... [et al.]
Volume60
StatePublished - 2013
Externally publishedYes

Fingerprint

Photobleaching
Optical Imaging
Endoplasmic Reticulum
Membrane Proteins
Proteins
Unfolded Protein Response
Protein Unfolding
Endoplasmic Reticulum Stress
Post Translational Protein Processing
Heat-Shock Proteins
Fluorescence Microscopy
Research Personnel
Calcium
Lipids
Membranes
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Medicine(all)

Cite this

@article{bb1d150c107f4abe9d3baaa8da2611f3,
title = "Probing endoplasmic reticulum dynamics using fluorescence imaging and photobleaching techniques.",
abstract = "This unit describes approaches and tools for studying the dynamics and organization of endoplasmic reticulum (ER) membranes and proteins in living cells using fluorescence microscopy. The ER plays a key role in secretory protein biogenesis, calcium regulation, and lipid synthesis. However, study of these processes has often been restricted to biochemical assays that average millions of lysed cells or imaging of static fixed cells. With new fluorescent protein (FP) reporter tools, sensitive commercial microscopes, and photobleaching techniques, investigators can interrogate the behaviors of ER proteins, membranes, and stress pathways in single live cells. Solutions are described for imaging challenges relevant to the ER, including the mobility of ER membranes, a range of ER structures, and the influence of post-translational modifications on FP reporters. Considerations for performing photobleaching assays for ER proteins are discussed. Finally, reporters and drugs for studying misfolded secretory protein stress and the unfolded protein response are described.",
author = "Lindsey Costantini and Erik Snapp",
year = "2013",
language = "English (US)",
volume = "60",
journal = "Current protocols in cell biology / editorial board, Juan S. Bonifacino ... [et al.]",
issn = "1934-2500",
publisher = "John Wiley and Sons Inc.",

}

TY - JOUR

T1 - Probing endoplasmic reticulum dynamics using fluorescence imaging and photobleaching techniques.

AU - Costantini, Lindsey

AU - Snapp, Erik

PY - 2013

Y1 - 2013

N2 - This unit describes approaches and tools for studying the dynamics and organization of endoplasmic reticulum (ER) membranes and proteins in living cells using fluorescence microscopy. The ER plays a key role in secretory protein biogenesis, calcium regulation, and lipid synthesis. However, study of these processes has often been restricted to biochemical assays that average millions of lysed cells or imaging of static fixed cells. With new fluorescent protein (FP) reporter tools, sensitive commercial microscopes, and photobleaching techniques, investigators can interrogate the behaviors of ER proteins, membranes, and stress pathways in single live cells. Solutions are described for imaging challenges relevant to the ER, including the mobility of ER membranes, a range of ER structures, and the influence of post-translational modifications on FP reporters. Considerations for performing photobleaching assays for ER proteins are discussed. Finally, reporters and drugs for studying misfolded secretory protein stress and the unfolded protein response are described.

AB - This unit describes approaches and tools for studying the dynamics and organization of endoplasmic reticulum (ER) membranes and proteins in living cells using fluorescence microscopy. The ER plays a key role in secretory protein biogenesis, calcium regulation, and lipid synthesis. However, study of these processes has often been restricted to biochemical assays that average millions of lysed cells or imaging of static fixed cells. With new fluorescent protein (FP) reporter tools, sensitive commercial microscopes, and photobleaching techniques, investigators can interrogate the behaviors of ER proteins, membranes, and stress pathways in single live cells. Solutions are described for imaging challenges relevant to the ER, including the mobility of ER membranes, a range of ER structures, and the influence of post-translational modifications on FP reporters. Considerations for performing photobleaching assays for ER proteins are discussed. Finally, reporters and drugs for studying misfolded secretory protein stress and the unfolded protein response are described.

UR - http://www.scopus.com/inward/record.url?scp=84907401235&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84907401235&partnerID=8YFLogxK

M3 - Article

C2 - 24510787

VL - 60

JO - Current protocols in cell biology / editorial board, Juan S. Bonifacino ... [et al.]

JF - Current protocols in cell biology / editorial board, Juan S. Bonifacino ... [et al.]

SN - 1934-2500

ER -