Methods for making and observing model lipid droplets

被引：4

作者：

Gandhi, Sonali A. ^{[1
]}

Parveen, Shahnaz ^{[1
]}

Alduhailan, Munirah ^{[1
]}

Tripathi, Ramesh ^{[1
]}

Junedi, Nasser ^{[1
]}

Saqallah, Mohammad ^{[1
]}

Sanders, Matthew A. ^{[2
,5
]}

Hoffmann, Peter M. ^{[1
,3
]}

Truex, Katherine ^{[4
]}

Granneman, James G. ^{[2
,5
]}

Kelly, Christopher, V ^{[1
,5
]}

机构：

[1] Wayne State Univ, Dept Phys & Astron, Detroit, MI 48201 USA

[2] Wayne State Univ, Ctr Mol Med & Genet, Sch Med, Detroit, MI 40201 USA

[3] Embry Riddle Aeronaut Univ, Phys Sci Dept, Daytona Beach, FL 32114 USA

[4] US Naval Acad, Dept Phys, Annapolis, MD 21402 USA

[5] Wayne State Univ, Ctr Integrat Metab & Endocrine Res, Sch Med, Detroit, MI 48201 USA

来源：

CELL REPORTS METHODS | 2024年 / 4卷 / 05期

基金：

美国国家科学基金会; 美国国家卫生研究院;

关键词：

SURFACE-TENSION; FAT; LIPOLYSIS; PROBES; ABHD5;

D O I：

10.1016/j.crmeth.2024.100774

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We present methods for making and testing the membrane biophysics of model lipid droplets (LDs). Methods are described for imaging LDs ranging in size from 0.1 to 40 mm in diameter with high-resolution microscopy and spectroscopy. With known LD compositions, membrane binding, sorting, diffusion, and tension were measured via fluorescence correlation spectroscopy (FCS), fluorescence recovery after photobleaching (FRAP), fluorescence lifetime imaging microscopy (FLIM), atomic force microscopy (AFM), and imaging flow cytometry. Additionally, a custom, small-volume pendant droplet tensiometer is described and used to measure the association of phospholipids to the LD surface. These complementary, cross-validating methods of measuring LD membrane behavior reveal the interplay of biophysical processes on lipid droplet monolayers.

引用

页数：19

共 50 条

[1] Protocol Making Droplet-Embedded Vesicles to Model Cellular Lipid Droplets
Chorlay, Aymeric
Santinho, Alexandre
Thiam, Abdou Rachid
STAR PROTOCOLS, 2020, 1 (03):
[2] Methods in Cell Biology Lipid Droplets Preface
Yang, Hongyuan
Li, Peng
LIPID DROPLETS, 2013, 116 : XIII - XIII
[3] Nanoemulsions of lipid droplets - a new model lipid system
Vezocnik, V.
Hodnik, V.
Okur, H. I.
Sitar, S.
Tusek-Znidaric, M.
Kogej, K.
Sepcic, K.
Roke, S.
Zagar, E.
Macek, P.
FEBS OPEN BIO, 2018, 8 : 473 - 473
[4] Fixation methods for the study of lipid droplets by immunofluorescence microscopy
DiDonato, D
Brasaemle, DL
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 2003, 51 (06) : 773 - 780
[5] Making and breaking Lipid Droplets : The Rough and the Smooth (ER) of it.
Mallik, R.
Kumar, M.
Ojha, S.
Joshi, A.
Kamat, S. S.
MOLECULAR BIOLOGY OF THE CELL, 2018, 29 (26)
[6] Phospholipids diffusion on the Surface of Model Lipid Droplets
Asfia, Shima
Seemann, Ralf
Fleury, Jean-Baptiste
EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS, 2023, 52 (SUPPL 1): : S116 - S116
[7] Phospholipids diffusion on the surface of model lipid droplets
Asfia, Shima
Seemann, Ralf
Fleury, Jean-Baptiste
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, 2023, 1865 (01):
[8] Model for Lipid Droplets within Endoplasmic Reticulum
Golani, Gonen
Kozlov, Michael M.
BIOPHYSICAL JOURNAL, 2017, 112 (03) : 83A - 83A
[9] A Model of Lipid Monolayer-Bilayer Fusion of Lipid Droplets and Peroxisomes
Kalutsky, Maksim A.
Galimzyanov, Timur R.
Molotkovsky, Rodion J.
MEMBRANES, 2022, 12 (10)
[10] Optimized methods to image hepatic lipid droplets in zebrafish larvae
Khan, Nouf
Salmi, Talhah Mohd
Karamalakis, Anthony P.
Nair, Anjana Ramdas
Sadler, Kirsten C.
Cox, Andrew G.
DISEASE MODELS & MECHANISMS, 2024, 17 (11)

← 1 2 3 4 5 →