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TIRF Imaging
TIRF Objectives and IXplore TIRF

High-Resolution Objectives for TIRF

A high numerical aperture (NA) is important for TIRF microscopy. As a pioneer in TIRF microscopy, we offer a diverse lineup of objectives with a high NA, ranging from 1.45 to the world’s highest NA of 1.7*1 and from 60x to 150x magnification. Modern observation methods, like super resolution and capturing images over a large field of view with an sCMOS camera, demand the highest quality objectives. That’s why we developed advanced objective manufacturing technology. This advanced polishing technology enabled us to create the world’s first plan-corrected apochromat objectives with an NA of 1.5*2. These objectives deliver uniform image quality, even over a large field of view, and are ideal for TIRF imaging.

*1 As of Nov, 2018. According to Olympus research.
*2 As of Nov, 2018. According to Olympus research.

Total Internal Reflection Fluorescence (TIRF) Objectives

Time–Lapse TIRF Imaging of Polymerization and Depolymerization Between a Plasma Membrane and FBP17 Membrane–Bending Protein

The penetration depth was adjusted to get a high signal-to-noise ratio with a high numerical aperture. 
TIRF imaging enables new types of advanced cellular research. This show changes in the membrane morphology and molecular dynamics under the cell membrane. The images were acquired using the UAPON100XOTIRF objective.*4
*4 Previous model of the UPLAP0100XOHR

Image data courtesy of Kazuya Tsujita, Ph.D., Toshiki Itoh, Ph.D., Biosignal Research Center, Organization of Advanced Science and Technology, Kobe University Reference: Nat Cell Biol. 2015 Jun; 17 (6): 749–58. doi: 10.1038/ ncb3162.

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Single–Molecule Fluorescence Imaging to Count the Subunits of a Transmembrane Ion Channel Complex (APON100XHOTIRF)

The objective enables single-molecule TIRF imaging with higher resolution and brighter images thanks to a numerical aperture of 1.70.

The subunit counting*5 technique was used to analyze the number of accessory dipeptidyl peptidase-like protein 10 (DPP10) molecules, which bind to the transmembrane ion channel Kv4.2, in one Kv4.2–DPP10 complex. The APON100XHOTIRF objective’s high NA enables researchers to measure fluorescence intensity change caused by photobleaching of a single molecule. This study*revealed that a maximum of 4 molecules of DPP10 subunits form a complex with the ion channel Kv4.2.

*5 Ulbrich, MH, and Isacoff EY. “Subunit counting in membrane–bound proteins.” Nature Methods, 4 (2007): 319–321.
*6 Kitazawa M, Kubo Y, and Nakajo K. “Kv4.2 and accessory dipeptidyl peptidase–like protein 10 (DPP10) subunit preferentially form a 4:2 (Kv4.2:DPP10) channel complex.” J Biol Chem, 290 (2015): 22724–22733.

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Determining the Subunit Stoichiometry of the Kv4.2–DPP10 Channel Complex by Subunit Counting

Schematic illustration of subunit counting of a transmembrane ion channel complex using single–molecule fluorescence imaging

TIRF Objectives Selection Guide

Working Distance
(mm)
Magnification Objective Field Number*3 Numerical Aperture Immersion Applications
UPLAPO60XOHR 0.11 60X 22 1.50 Oil Real-time, super resolution imaging for live cells/super resolution imaging of tiny structures, such as organelles/whole-cell TIRF imaging
UPLAPO100XOHR 0.12 100X 22 1.50 Oil Real-time, super resolution imaging for live cells/super resolution imaging of tiny structures, such as organelles/high-resolution imaging of cell membranes or subcellular organelles, and single-molecule-level experiments
APON100XHOTIRF 0.08 100X 22 1.70 Special Oil Observing the movement of proteins or vesicles at the single-molecule level
UAPON150XOTIRF 0.08 150X 22 1.45 Oil Subcellular imaging (such as organelles, endoplasmic reticulum, and intracellular vesicle trafficking) 

*3 Maximum field number observable through eyepiece

Related Products

IXplore TIRF

IXplore TIRF

  • Utilize the Olympus real-time controller for physiologically relevant data with minimal cell disturbance 
  • Maintain cell viability while imaging with various environmental control options 
  • Maintain focus accurately and reliably in time-lapse experiments with the Olympus hardware autofocus (Z-drift compensation) system 
  • Discover the real shape of your cells with Olympus silicone immersion optics 

Learn More

*Banner Image: By courtesy of Dr. Michael W. Davidson The Florida State University

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