Cost-efficient nanoscopy reveals nanoscale architecture of liver cells and platelets
Mao Hong,
Diekmann Robin,
Liang Hai Po H.,
Cogger Victoria C.,
Le Couteur David G.,
Lockwood Glen P.,
Hunt Nicholas J.,
Schüttpelz Mark,
Huser Thomas R.,
Chen Vivien M.,
McCourt Peter A.G.
Affiliations
Mao Hong
Faculty of Health Sciences, Department of Medical Biology, University of Tromsø-The Arctic University of Norway, Hansine Hansens veg 18, Tromsø 9037, Norway, e-mail: [email protected]
Diekmann Robin
EMBL Heidelberg, Cell Biology and Biophysics Unit, 69117 Heidelberg, Germany
Liang Hai Po H.
ANZAC Research Institute, Concord Repatriation General Hospital, Concord, NSW, Australia
Cogger Victoria C.
ANZAC Research Institute, Concord Repatriation General Hospital, Concord, NSW, Australia
Le Couteur David G.
ANZAC Research Institute, Concord Repatriation General Hospital, Concord, NSW, Australia
Lockwood Glen P.
ANZAC Research Institute, Concord Repatriation General Hospital, Concord, NSW, Australia
Hunt Nicholas J.
ANZAC Research Institute, Concord Repatriation General Hospital, Concord, NSW, Australia
Schüttpelz Mark
Department of Physics, Bielefeld University, 33615 Bielefeld, Germany
Huser Thomas R.
Department of Physics, Bielefeld University, 33615 Bielefeld, Germany
Chen Vivien M.
ANZAC Research Institute, Concord Repatriation General Hospital, Concord, NSW, Australia
McCourt Peter A.G.
Department of Medical Biology, University of Tromsø-The Arctic University of Norway, 9037 Tromsø, Norway
Single-molecule localization microscopy (SMLM) provides a powerful toolkit to specifically resolve intracellular structures on the nanometer scale, even approaching resolution classically reserved for electron microscopy (EM). Although instruments for SMLM are technically simple to implement, researchers tend to stick to commercial microscopes for SMLM implementations. Here we report the construction and use of a “custom-built” multi-color channel SMLM system to study liver sinusoidal endothelial cells (LSECs) and platelets, which costs significantly less than a commercial system. This microscope allows the introduction of highly affordable and low-maintenance SMLM hardware and methods to laboratories that, for example, lack access to core facilities housing high-end commercial microscopes for SMLM and EM. Using our custom-built microscope and freely available software from image acquisition to analysis, we image LSECs and platelets with lateral resolution down to about 50 nm. Furthermore, we use this microscope to examine the effect of drugs and toxins on cellular morphology.
