Quad band pass dichroic filter

Manufactured by Nikon

The Quad-band-pass dichroic filter is a specialized optical component designed to selectively transmit and reflect specific wavelength bands. It consists of multiple dielectric coatings that allow the passage of four distinct wavelength ranges while reflecting all other wavelengths. This filter is a key element in various laboratory and research applications that require precise control and separation of light.

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Lab products found in correlation

N storm system, by Nikon (2 mentions) N storm 4.0 system, by Nikon (2 mentions) Orca flash 4.0 cmos camera, by Hamamatsu Photonics (1 mentions) Glass slide, by Corning (1 mentions) Phalloidin 647, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 anti rat, by Thermo Fisher Scientific (1 mentions) Scmos camera, by Hamamatsu Photonics (1 mentions) Tubulin rat antibody, by Abcam (1 mentions) Anti mouse alexa fluor 555, by Thermo Fisher Scientific (1 mentions)

6 protocols using quad band pass dichroic filter

Super-Resolution Imaging of Fluorescent Nanoparticles

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dSTORM imaging was
performed with a Nikon
N-STORM system configured for TIRF imaging and equipped with a perfect
focus system. AF647-labeled proteins were illuminated using a 647
nm laser (170 mW), and sicastar-greenF NPs were illuminated using
a 488 nm laser (90 mW) with an adjusted TIRF angle to maximize the
signal-to-noise ratio. No UV activation was used. A Nikon 100X, 1.4
NA oil immersion objective was used to collect the fluorescence signal,
which was passed through a quad-band-pass dichroic filter (97335,
Nikon) and recorded on an Andor EMCCD camera (ixon3) with pixel size
160 nm and a region of interest of 256 × 256 pixels.

Woythe L., Madhikar P., Feiner-Gracia N., Storm C, & Albertazzi L. (2022). A Single-Molecule View at Nanoparticle Targeting Selectivity: Correlating Ligand Functionality and Cell Receptor Density. ACS Nano, 16(3), 3785-3796.

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Super-Resolution Imaging of BTA Fibers

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STORM imaging was performed as previously described16 (link). BTA fibres were immobilized by adsorption onto the surface of a flow chamber assembled from a glass slide and a coverslip separated by double-sided tape. Glass microscope coverslips (thickness 0.17 mm) were washed as previously decribed16 (link). Images were acquired using a Nikon N-STORM system configured for TIRF imaging. Cy5-labelled samples were illuminated by the 647-nm laser lines built into the microscope. Fluorescence was collected by means of a Nikon × 100, 1.4 NA oil immersion objective and passed through a quad-band-pass dichroic filter (97335 Nikon). Frames (30,000) were recorded onto a 128 × 128 pixel region of an EMCCD camera (ixon3, Andor). STORM images are reconstructed with the STORM module of the NIS element Nikon software.

Baker M.B., Albertazzi L., Voets I.K., Leenders C.M., Palmans A.R., Pavan G.M, & Meijer E.W. (2015). Consequences of chirality on the dynamics of a water-soluble supramolecular polymer. Nature Communications, 6, 6234.

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Visualization of Cy3-Labeled BTA Fibers

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TIRF images were
acquired with a Nikon N-STORM system. Cy3 was excited
using a 561 nm laser. Fluorescence was collected by means of a Nikon
×100, 1.4NA oil immersion objective and passed through a quad-band
pass dichroic filter (97335 Nikon). Images were recorded with an EMCCD
camera (ixon3, Andor, pixel size 0.17 μm). BTA fibers were prepared
at a total concentration of 20 μM containing Cy3-labeled BTA-OEG₄ (Cy3-BTA-OEG₄)
and further diluted to 2.5 μM and flown in a chamber between
a glass microscope coverslip (Menzel-Gläser, no. 1.21 ×
26 mm) and a glass slide, which were separated by double-sided tape.

Varela-Aramburu S., Su L., Mosquera J., Morgese G., Schoenmakers S.M., Cardinaels R., Palmans A.R, & Meijer E.W. (2021). Introducing Hyaluronic Acid into Supramolecular Polymers and Hydrogels. Biomacromolecules, 22(11), 4633-4641.

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Nanoparticle characterization by TIRF

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30 μL of 10-fold diluted particles suspension was introduced into a flow chamber created with 24x24 mm glass cover slip (RS, France) attached with a two face Scotch tape to the edges of a glass slide (25x75 mm, Corning). The sample was incubated for 15 min. at RT and next 100 μL of purified water were introduced into the flow chamber to flush away unattached NPs.
Images were acquired using a Nikon N-STORM 4.0 system conFigd for total internal reflection (TIR) fluorescence, using a Perfect Focus System imaging. Excitation under the TIR conditions allowed to avoid illumination of out of focus, improving signal to noise ratio. DiI fluorophore was excited by illuminating the sample with a 5% power of 561 nm (80 mW) laser built into the microscope. During acquisition the integration time was 300 ms. Fluorescence was collected by means of a Nikon x100, 1.4 NA oil immersion objective and passed through a quad-band-pass dichroic filter (97335 Nikon). Images were recorded onto a 256 x 256-pixel region (pixel size 160 nm) of a Hamamatsu ORCA Flash 4.0 CMOS camera. The images were analyzed using ImageJ software. Briefly, the intensity threshold was set to filter the NPs in each image, and next the fluorescence intensity per particle was measured (for minimum 800 NPs) and plotted in a histogram graph.

Mares A.G., Pacassoni G., Marti J.S., Pujals S, & Albertazzi L. (2021). Formulation of tunable size PLGA-PEG nanoparticles for drug delivery using microfluidic technology. PLoS ONE, 16(6), e0251821.

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Two-color iPAINT Imaging Protocol

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iPAINT images are acquired using a Nikon
N-STORM system equipped with ∼160 mW/cm² (λ
= 405 nm), ∼488 mW/cm² (λ = 561 nm), and ∼1.3
W/cm² (λ = 647 nm) laser lines configured for quasi-total internal reflection fluorescence imaging. The
angle at which the inclined excitation is performed is finely tuned
to maximize the signal-to-noise ratio. The fluorescence emission is
collected by means of an oil immersion objective (Nikon 100×,
1.49 NA). A quad-band-pass dichroic filter (97335 Nikon) is used to
separate the excitation light from the fluorescence emission. Time-lapse
images of 15 × 10³ frames are recorded onto a 256
× 256 pixel region (pixel size 170 nm) of an EMCCD camera (ixon3,
Andor) at a rate of 47 frames/s. Two different photoactivatable dyes
are used to perform two-color iPAINT measurements: Cage-635 and Cage-552
(Abberior) (Scheme S1a,b, respectively).
Upon excitation with the proper wavelength (λ = 561 nm for Cage-552,
λ = 647 nm for Cage-635), the fluorescence of the dyes is collected.
The localization of single molecules in the iPAINT image is carried
out by NIS-element Nikon software.

Adelizzi B., Aloi A., Van Zee N.J., Palmans A.R., Meijer E.W, & Voets I.K. (2018). Painting Supramolecular Polymers in Organic Solvents by Super-resolution Microscopy. ACS Nano, 12(5), 4431-4439.

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STORM Imaging of Cardiac F-Actin

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For STORM imaging, hearts were seeded on glass petri dishes, and F-actin was labeled with phalloidin-647 (ThermoFisher, A22287) at 1:1000 dilution in PBS. Additional immunostaining was performed using the following primary antibodies: Paxillin mouse antibody (BD Biosciences, 610051) and tubulin rat antibody (Abcam, ab6160). Additional secondary antibodies were: Alexa Fluor 488 anti-rat (ThermoFisher, A11006) and Alexa Fluor 555 anti-mouse (ThermoFisher, A21424). All antibodies were diluted 1:1000.
Images were acquired using a Nikon N-STORM 4.0 system configured for total internal reflection fluorescence (TIRF) imaging. Excitation inclination was tuned to adjust focus and to maximize the signal-to-noise ratio. For STORM imaging Alexa647 was excited illuminating the sample with the 647 nm (∼160 mW) laser line built into the microscope. Fluorescence was collected by means of a Nikon ×100, 1.4 NA oil immersion objective and passed through a quad-band-pass dichroic filter (97335 Nikon). Images were recorded on a 256×256 pixel region (pixel size 160 nm) of a sCMOS camera (Hamamatsu). Samples were kept in Gloxy buffer for STORM imaging as previously described 57 . Single-molecule localization sequences were analyzed with the STORM plug-in of NIS element Nikon software.

Uroz M., Garcia-Puig A., Tekeli I., Elosegui-Artola A., Abenza J.F., Marín-Llauradó A., Pujals S., Conte V., Albertazzi L., Roca-Cusachs P., Raya Á, & Trepat X. (2019). Traction forces at the cytokinetic ring regulate cell division and polyploidy in the migrating zebrafish epicardium. Nature materials, 18(9).

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