Ff02 520 28

Manufactured by IDEX Corporation

The FF02-520/28 is a filter module designed for use in laboratory applications. It features a 520 nm center wavelength and a 28 nm bandwidth. The product is designed to meet the needs of customers requiring optical filtering capabilities in their research or testing environments.

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Lab products found in correlation

Calcium calibration buffer kit 1, by Thermo Fisher Scientific (1 mentions) Fusion α, by PerkinElmer (1 mentions) 96 well plate, by PerkinElmer (1 mentions) Originpro 7, by OriginLab (1 mentions) Uplanapo, by Olympus (1 mentions) Orca fusion scmos camera, by Hamamatsu Photonics (1 mentions) W view gemini image splitter, by Hamamatsu Photonics (1 mentions)

3 protocols using ff02 520 28

Two-Photon Imaging of Oxytocinergic Neurons

Cited 1 time

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We recorded RB-positive or negative oxytocinergic neurons in the PVN with potassium-based internal solution containing Alexa Fluor 488. After 15–20 min-long recording, cell morphology was visualized using Alexa Fluor 488 (10 – 20 μM) excited with 910 nm light. The beam of an 80 MHz Ti:Sapphire laser (Mai Tai eHP DS, Newport) was directed by a two-dimensional galvanometer scanning mirror system (HSA Galvo 8315K, Cambridge Technology). Fluorescence emission was collected by two PMTs above and below the sample (H10770P, Hamamatsu) after passing through a dichroic beam splitter (FF670-SDi01–26×38, Semrock) and a bandpass filter (FF02–520/28, Semrock). We used ScanImage 3.8 to control scanning parameters and image acquisition of 1.5 μm-thick z-stacks through each recorded neuron (Pologruto et al., 2003 (link)). Laser intensity was controlled by a Pockels cell and laser power at the sample plane was 10–15 mW. Area of the soma was measured on z-projections with FIJI/ImageJ (Schindelin et al., 2012 (link)).

Xiao L., Priest M.F., Nasenbeny J., Lu T, & Kozorovitskiy Y. (2017). Biased oxytocinergic modulation of midbrain dopamine systems. Neuron, 95(2), 368-384.e5.

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Fluorometric calcium indicator calibration

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In vitro fluorometry was performed in 96-well plates (PerkinElmer) using a Fusion α plate reader (PerkinElmer) at room temperature. Filter sets used for fluorescent measurement for green calcium indicators were ET495/10x (Chroma) (excitation) and FF02-520/28 (Semrock) (emission). The dynamic range was calculated as ΔF/F. ΔF/F was calculated as (F_max–F₀)/F₀, where F_max is the fluorescence intensity at a saturating [Ca²⁺] of 0.3 mM and F₀ is the fluorescence intensity measured at zero [Ca²⁺] in the presence of 15 mM EGTA. Calcium titration experiments were performed by mixing 10 mM solutions of K₂H₂EGTA and Ca₂EGTA proportionally from Calcium Calibration Buffer Kit #1 (Thermo Fisher Scientific). The K_d value and Hill coefficient were calculated by fitting according to the Hill equation using Origin Pro 7.5 (Origin Lab).

Sakamoto M., Inoue M., Takeuchi A., Kobari S., Yokoyama T., Horigane S.I., Takemoto-Kimura S., Abe M., Sakimura K., Kano M., Kitamura K., Fujii H, & Bito H. (2022). A Flp-dependent G-CaMP9a transgenic mouse for neuronal imaging in vivo. Cell Reports Methods, 2(2), 100168.

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Multicolor TIRF Microscopy Imaging

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All TIRF imaging data was collected using an inverted microscope (IX83; Olympus) equipped with a 60×/1.5 NA oil-immersion TIRF (UPlanApo; Olympus) and included a correction collar, with an additional 1.6X magnification applied (total 100X magnification). Fluorescent excitation used a 640 nm/140 mW diode laser (Olympus Soft Imaging Solution; 00026121) and a 488 nm/100mW diode laser (Olympus Soft Imaging Solution; 00026125). Images were captured using a Hamamatsu ORCA-Fusion sCMOS camera (C14440) and Hamamatsu W-View Gemini image splitter (A12801-01). For simultaneous 2-color, live cell acquisition a filter set of Chroma single-bandpass 705/100 nm (394148) and a Semrock 520/28 nm (FF02-520/28) were used in combination with a Semrock single-edge standard epi-fluorescence dichroic beamsplitter (FF555-Di03). Images were acquired at 900 ms exposure time. Samples were held at 36°C using a Bioptechs objective heater controller (150803).

Rinaldi D.A., Kanagy W.K., Kaye H.C., Grattan R.M., Lucero S.R., Pérez M.P., Wester M.J., Lidke K.A., Wilson B.S, & Lidke D.S. (2023). Antigen Geometry Tunes Mast Cell Signaling Through Distinct FcεRI Aggregation and Structural Changes. bioRxiv.

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