Ff495 di03

Manufactured by IDEX Corporation

The FF495-Di03 is a laboratory equipment product manufactured by IDEX Corporation. It serves as a filter, designed to isolate specific wavelengths of light during scientific experiments and analyses. The core function of this device is to selectively transmit or block particular wavelengths of light, enabling researchers to control the spectral composition of the light being used in their experiments.

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Close spelling variants of this product

FF495-Di03-25 × 36 (4 citations) FF495 (3 citations) FF495-Di03-50.8-D dichroic mirror (3 citations) FF495-Di03-25x36 (2 citations)

6 protocols using ff495 di03

3D Holographic Tracking of Sperm Cells

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Freely swimming sperm were tracked using an inverted microscope (IX71; Objective × 20, 0.75 numerical aperture, UPLSAPO; Olympus). Coherent illumination was achieved with a laser light source (LDH-D-C-510, PicoQuant GmbH) and the corresponding controller (Sepia II Multichannel Processor, PicoQuant GmbH). Laser light was coupled into a multimode fibre. A custom-made adapter was used to position the fibre parallel to the optical axis of the objective. The illumination intensity was adjusted to use the dynamical range of the camera (12-bit; PCO Dimax HD). Movies were collected at 600 frames per second; each frame represents a holographic image containing the complete 3D information of the sperm cell.
Caged compounds were photolyzed using a 365-nm LED (M365L2-C; Thorlabs). The ultraviolet light was coupled into a liquid guide (77566; Newport) followed by two Plano-convex lenses (LA 1951-A f=25.4 mm; LA 1509-A f=100 mm; Thorlabs) and coupled to the imaging optical path with a dichroic filter (ff 495-Di03; Semrock). The irradiation power (0.8 mW) was measured with a power meter (detector PowerMax and head model PS19Q; Coherent). The light spectrum was recorded with a spectrometer (51024 DW; Ocean Optics). Photolysis and data acquisition were synchronized using a wave generator (33500B; Agilent).

Jikeli J.F., Alvarez L., Friedrich B.M., Wilson L.G., Pascal R., Colin R., Pichlo M., Rennhack A., Brenker C, & Kaupp U.B. (2015). Sperm navigation along helical paths in 3D chemoattractant landscapes. Nature Communications, 6, 7985.

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High-Speed Fluorescence Imaging of ArcLight

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During the patch clamp experiments the cells were imaged on an IX71 microscope with a 60× 1.35-numerical aperture (NA) oil-immersion lens (Olympus). Fluorescence excitation was delivered using a 75 W Xenon Arc lamp (Cairn). The filter set used was that reported for ArcLight [7 (link)]. The excitation filter was FF02-472/30 (Semrock). The emission filter was FF01-496/LP (Semrock). The dichroic was FF495-Di03 (Semrock). The objective C-mount image was de-magnified by an Optem zoom system A45699 (Qioptiq LINOS, Inc., Fairport, NY) and projected onto the e2v CCD39 chip of NeuroCCD-SM 80 pixel × 80 pixel camera (RedShirtImaging,). The imaging apparatus was mounted on a Vibraplane Bench Top vibration isolation platform (Minus K Technology). The mechanical shutter in the incident light patch was mounted on a separate table. Images were recorded at a frame rate of 1kfps. The excitation light was 1 mW/mm².

Jung A., Rajakumar D., Yoon B.J, & Baker B.J. (2017). Modulating the Voltage-sensitivity of a Genetically Encoded Voltage Indicator. Experimental Neurobiology, 26(5), 241-251.

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High-Resolution Fluorescence Microscopy Imaging

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Images of HEK293 cells or neurons were acquired with an inverted microscope (IX71, Olympus) equipped with a 100×/1.45-NA oil-immersion objective. Cells were illuminated with high-power light-emitting diodes with wavelength of 405 nm (M405LP1, Thorlabs), 450 nm (M450LP1, Thorlabs) and/or 470 nm (M470 L3, Thorlabs). 405-nm and 470-nm illumination light were filtered at the exit of LEDs with band-pass filters centered at 405 nm (FBH405–10, Thorlabs) and 475 nm (FF01–475/28, Semrock), respectively. 450 nm illumination was filtered with a band-pass filters centered at 434 nm to allow the 440-nm component passing through. The fluorescence was collected through a 495-nm dichroic mirror (FF495-Di03, Semrock) and a 535/50 emission filter (FF01–535/50, Semrock). Illumination intensity was 5 mW/mm² in all cases for 470-nm light and and 0.1~ 2.5 mW/mm² for 405-nm light at the specimen plane. In all experiments, images were acquired continuously unless described explicitly, at 200 Hz with a sCMOS camera (Zyla, Andor). The camera and the microscope were connected with a 0.35×-magnification adapter (Olympus). Synchronization of electrophysiology and imaging was implemented with a DAQ board (PCI-6229, National Instruments) interfaced with Igor Pro (Wavemetrics) and Micro-Manager [20 (link)].

Xu F., Shi D.Q., Lau P.M., Lin M.Z, & Bi G.Q. (2018). Excitation wavelength optimization improves photostability of ASAP-family GEVIs. Molecular Brain, 11, 32.

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Customized LED Setup for Wavelength-Specific Stimulation

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We produced full-field wavelength-specific stimuli using a customized setup (Figure 1C). The setup consists of six LEDs in the UV and visible wavelength range (ThorLabs M340L4 - dUV/340nm; M365L2 - UV/360nm; M415L4 - violet/415nm; M455L3 - blue/455nm; M565L3 - lime/565nm; M617L3 - orange/615nm). A customized driver drove the five LEDs from dUV to lime. These LEDs turned on during the return period of the x-scanning mirror in the two-photon microscope (fly-back stimulation). We used the TTL signal generated by the two-photon microscope at the beginning of each line-scan of the horizontal scanning mirror (x-mirror) to trigger the LED driver. An individual T-Cube (Thorlabs LEDD1B T-Cube) drove the orange LED. Stimuli were generated using customized software written in Python. The update rate for the LED voltage values was 180Hz.
The different light sources were focused with an aspheric condenser lens (ThorLabs ACL2520U-A) and aligned using dichroic mirrors (dUV-UV dichroic - Semrock LPD01-355RU; UV-violet dichroic - Semrock FF414-Di01; violet-blue dichroic - Semrock Di02-R442; blue-lime dichroic - Semrock FF495-Di03; lime-orange dichroic - Semrock FF605-Di02). The collimated light passed through a diffuser (ThorLabs DG10-1500A) before reaching the eye of the fly, which is positioned 2cm away.

Heath S.L., Christenson M.P., Oriol E., Saavedra-Weisenhaus M., Kohn J.R, & Behnia R. (2020). Circuit mechanisms underlying chromatic encoding in Drosophila photoreceptors. Current biology : CB, 30(2), 264-275.e8.

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Cytosolic pH Measurement in HeLa Cells

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The cytosolic pH of HeLa cells was measured with BCECF (2′-7′-bis(carboxyethyl)-5(6)-carboxyfluorescein) as the following protocol. The cells were treated with 10 μM BCECF acetoxymethyl ester (Molecular probes) in Hank’s balanced salt solution (HBSS) for 15 min at 37 °C, followed by washing with HBSS twice. Fluorescence intensity of BCECF of the cells (n = 6) was measured at 37 °C in DMEM using a Nikon TE2000 (Nikon Instruments) equipped a CFI Plan Apo VC 60x oil-immersion objective lens, 1.40 numerical aperture (Nikon Instruments). The following filters (Semrock) were used for the measurement; excitation filters (FF01-445/20 and FF01-482/18), an emission filter (FF01-520/35), and a dichroic mirror (FF495-Di03). Images were captured with an ORCA-AG cooled CCD camera (Hamamatsu photonics). Calibration curve was obtained by imaging BCECF-loaded cells in pH-controlled buffers containing 150 mM KCl, 20 mM NaCl, 0.5 mM MgCl₂, 0.5 mM CaCl₂, 10 mM MES, 10 mM HEPES, 10 μM monensin and 10 μM nigericin.

Yoshida T., Kakizuka A, & Imamura H. (2016). BTeam, a Novel BRET-based Biosensor for the Accurate Quantification of ATP Concentration within Living Cells. Scientific Reports, 6, 39618.

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Wide-field Fluorescence Macroscope for Cortical Ca2+ Imaging

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To image neural Ca 2+ activity across 11 mouse cortical areas, we designed and built a custom wide-field fluorescence macroscope with a field-of-view spanning 4 mm in diameter (Fig. 1a). For epi-fluorescence illumination we used a light-emitting diode (LED) (Thorlabs M470L2) with an emission spectrum centered in the 440-480 nm range. The imaging pathway comprised an objective lens (Leica, 5.0× Planapo 0.5 NA; 19 mm working distance; anti-reflection coated for 400-1000 nm light; transmission >90% at 520 nm), a tube lens (75 mm focal length; Thorlabs AC508-075-A-ML), a custom fluorescence filter cube (excitation filter: Semrock FF01-466/40-25; dichroic mirror: Semrock FF495-Di03, custom-sized to 35 mm × 50 mm; emission filter: Semrock FF02-525/40, custom-sized to 30 mm × 30 mm), and a scientific-grade CMOS camera (Hamamatsu ORCA-Flash4.0 V2 sCMOS). To control image acquisition, we used HCImage software (Hamamatsu), which communicated with the camera via an Active Silicon Firebird Camera Link Board.

Ebrahimi S., Lecoq J., Rumyantsev O., Tasci T., Zhang Y., Irimia C., Li J., Ganguli S, & Schnitzer M.J. (2022). Emergent reliability in sensory cortical coding and inter-area communication. Nature, 605(7911).

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