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OMX BLAZE 3D-structured illumination, super-resolution microscope (SIM)

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RESERVE THIS MICROSCOPE

This microscope can achieve both lateral (XY) and axial (Z) resolution at twice the diffraction limit of conventional high-resolution light microscope systems (e.g. widefield deconvolution, confocal, 2-photon). The OMX BLAZE is capable of simultaneous, fast, multi-channel 3D SIM super-resolution imaging (tens of microns deep). In addition to the 3D SIM modality, the OMX BLAZE has also been designed for stochastic localization super resolution (STORM). Additionally, the optical paths for each excitation and emission wave length are separate and are engineered with dedicated, sensitive detectors (emCCDs). These unique features of the OMX BLAZE enable ultrafast live-cell 3D, simultaneous widefield imaging at diffraction-limited resolution with extremely high temporal resolution and rapid, multichannel 3D structured illumination super resolution. Additionally the OMX supports live and fixed sample 3D deconvolution.

System specifications

OMX V4 microscope platform with X, Y, Z Nanomotion stage movers with 25 mm travel

OMX BLAZE SI patterns generated using an electro-optical high-speed SI diffraction grating engine

Piezo controlled Fast Z-axis system - 100 micron range

Focus maintenance - Ultimate Focus System (UFS)

TIRF illumination module with 60X 1.49 TIRF lens

MONET/DLM for localization-type (aka PALM, STORM) super resolution. Note: AF647 is currently the only fluorophore that works for STORM on OMX.

60X, N.A. 1.30silicon immersion objective for matching of refractive indexes during live cell imaging

Environmental control (heating, CO2)

DIC optics

Off-line workstation with SoftWoRx software for generating SI image reconstructions

Objectives

Magnification

N. A.

Resolution x,y/z (nm)

Working Distance (um)

U-PLANAPO 100X SIM

1.40

??

150

ApoN TIRF 60x

1.49

??

100

U-ApoN 60X Silicone

1.3

??

150

Lasers

100mW: 405nm, 445nm, 488nm, 514nm, 568nm; 300 mW 642nm MONET laser

Detection

3 channels, emCCD cameras (Photometrics Evolve 512) cooled to -80oC back thinned frame transfer 512 x 512 pixels, pixel size, 16 um, readout speeds: 5 and 10 MHz.

Emission filters

DAPI or equivalent dye 435/31 nm

CFP or equivalent dye 477/35 nm

AlexaFluor488 or equivalent dye 528/48 nm

YFP or equivalent dye 541/22 nm

AlexaFluor568, mCherry or equivalent dye 609/37 nm

Cy5/AlexaFluor647 or equivalent dye 683/40 nm

Funding

This microscope was funded by a NIH High End Instrumentation grant (Dr. Aaron Straight PI). Installation date: 10/2012

Required Acknowledgement

The following acknowledgement is required when publishing data obtained with this instrument:"The project described was supported, in apart, by Award Number 1S10OD01227601 from the National Center for Research Resources (NCRR). Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the NCRR or the National Institutes of Health."

Location

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