easySTORM: a robust, lower-cost approach to localisation and TIRF microscopy.
TIRF and STORM microscopy are super-resolving fluorescence imaging modalities for which current implementations on standard microscopes can present significant complexity and cost. We present a straightforward and low-cost approach to implement STORM and TIRF taking advantage of multimode optical fibres and multimode diode lasers to provide the required excitation light. Combined with open source software and relatively simple protocols to prepare samples for STORM, including the use of Vectashield for non-TIRF imaging, this approach enables TIRF and STORM imaging of cells labelled with appropriate dyes or expressing suitable fluorescent proteins to become widely accessible at low cost.
Photonics Group, Physics Department, Imperial College London, London, SW7 2AZ. k.kwakwa@imperial.ac.uk.
Kwasi
K
Kwakwa
Photonics Group, Physics Department, Imperial College London, London, SW7 2AZ.|Institute of Chemical Biology, Imperial College London, London, SW7 2AZ.
Alexander
A
Savell
MRC Clinical Sciences Centre, Imperial College London, Du Cane Road, London, W12 0NN.
Timothy
T
Davies
Photonics Group, Physics Department, Imperial College London, London, SW7 2AZ.
Ian
I
Munro
MRC Clinical Sciences Centre, Imperial College London, Du Cane Road, London, W12 0NN.
Simona
S
Parrinello
Section of Hepatology, QEQM Hospital, Imperial College London, London, W2 1PG, UK.
Marco A
MA
Purbhoo
Photonics Group, Physics Department, Imperial College London, London, SW7 2AZ.|Centre for Pathology, Imperial College London, London, W12 0NN.
Chris
C
Dunsby
Photonics Group, Physics Department, Imperial College London, London, SW7 2AZ.
Mark A A
MA
Neil
Photonics Group, Physics Department, Imperial College London, London, SW7 2AZ.
Paul M W
PM
French
D000818:Animals; D045744:Cell Line, Tumor; D042783:Endothelial Cells / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology; D005456:Fluorescent Dyes; D006801:Humans; D007834:Lasers; D008027:Light; D051379:Mice; D008856:Microscopy, Fluorescence / Q000191:economics / Q000379:methods*; D055100:Optical Fibers; D011506:Proteins; D012984:Software
D005456:Fluorescent Dyes; D011506:Proteins
Microscopy; PALM; STORM; localization; super-resolution