Ff02 438 24

Manufactured by IDEX Corporation

The FF02-438/24 is a laboratory equipment product manufactured by IDEX Corporation. It serves as a core functional component for various scientific applications. The detailed specifications and intended uses are not included in this factual description.

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Lab products found in correlation

Ff01 650 13 25, by IDEX Corporation (1 mentions) Ff01 600 14 25, by IDEX Corporation (1 mentions) Mvplapo 1, by Olympus (1 mentions) Ff02 632 22 25, by IDEX Corporation (1 mentions) Ff01 692 40, by IDEX Corporation (1 mentions) Ti2 ctre microscope controller, by Nikon (1 mentions) Ti2 s se e motorized stage, by Nikon (1 mentions) Prime 95b, by Teledyne (1 mentions) Metamorph software, by Molecular Devices (1 mentions) Eclipse ti2, by Nikon (1 mentions) Pma 12 photonic multichannel analyzer, by Hamamatsu Photonics (1 mentions) Ff458 di02 25x36, by IDEX Corporation (1 mentions) Ixon ultra 897, by Oxford Instruments (1 mentions) Cfi plan apo vc 20, by Nikon (1 mentions) Ff01 542 27, by IDEX Corporation (1 mentions)

7 protocols using ff02 438 24

Automated Fluorescence Imaging Device

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We built a device equipped with an optics enabling automated acquisition of florescence images of a sample. A schematic view of the optical design is shown in Fig. 1. We placed the sample in a cassette, and the sample was illuminated by the light emitting from an LED unit. The unit consists of two tunable LEDs (CBT-90-UV and CBT-90-B, Luminus), which have excitation filters of either 405 nm or 435 nm (FF01-406/15, FF02-438/24, Semrock), respectively. Fluorescence emitting from the sample was reflected by the dichroic mirror at wavelengths longer than 470 nm, collimated with an objective lens (MVPLAPO × 0.63, Olympus), and imaged on a monochrome CCD camera (Lt665R, Lumenera). An optical filter unit was placed at the position between the objective lens and the imaging lens (MVPLAPO × 1, Olympus). The optical filter unit consisted of fluorescence filters with the center wavelengths of 600 nm, 632 nm, 650 nm and 680 nm (FF01-600/14-25, FF02-632/22-25, FF01-650/13-25, and FF01-680/22-25, Semrock).Fig. 1

Schematic view of the device for automated detection of LN metastasis

Matsumoto T., Murayama Y., Matsuo H., Okochi K., Koshiishi N., Harada Y., Tanaka H., Takamatsu T, & Otsuji E. (2020). 5-ALA-assistant automated detection of lymph node metastasis in gastric cancer patients. Gastric Cancer, 23(4), 725-733.

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Confocal Imaging of dsRNA Production

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For analysis of the production of dsRNA, confocal images were acquired with Eclipse Ti2 (Nikon) microscope, equipped with a PlanApo 20×/0.8 objective lens, a TI2-CTRE microscope controller (Nikon), a TI2-S-SE-E motorized stage (Nikon), an X-Light V3 (CrestOptics) spinning disk confocal unit, and a PRIME 95B scientific complementary metal-oxide semiconductor (sCMOS) camera (Teledyne Photometrics). The cells were illuminated with a CELESTA Light Engine laser light source (Lumencor) through a an FF01-391/477/549/639/741 excitation filter. Emission filters adopted for this experiment included an FF02-438/24 (Semrock) for Hoechst 33342, and an FF01-692/40 (Semrock) for Alexa 647. An FF421/491/567/659/776-Di01 dichroic mirror (Semrock) was used throughout this observation. Total fluorescent intensity within cells was quantified by using the ‘Multi Wavelength Cell Scoring’ module of MetaMorph software (Molecular Devices).

Tamura T., Torii S., Kajiwara K., Anzai I., Fujioka Y., Noda K., Taguwa S., Morioka Y., Suzuki R., Fauzyah Y., Ono C., Ohba Y., Okada M., Fukuhara T, & Matsuura Y. (2022). Secretory glycoprotein NS1 plays a crucial role in the particle formation of flaviviruses. PLoS Pathogens, 18(6), e1010593.

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Fluorescence and Luminescence Spectroscopy

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For the measurement of fluorescence spectra, HeLa cells expressing CFP alone, YFP alone, or a biosensor were plated onto a 35 mm glass-based dish. Cells were observed with an inverted microscope (IX81; Olympus, Tokyo) equipped with an objective lens (UPLAPO 100×/1.35NA oil objective; Olympus). CFP were excited by an FF02-438/24 (Semrock) excitation filter and an FF458-Di02-25x36 (Semrock) dichroic mirror. YFP were excited by an S492/18X (Chroma) excitation filter and a glass dichroic mirror (Olympus). Fluorescence spectra were recorded at 2 nm intervals by using a PMA-12 photonic multichannel analyzer (Hamamatsu Photonics, Hamamatsu, Japan). For the measurement of luminescent spectra, HeLa cells expressing a biosensor were trypsinized and suspended in M199 (ThermoFisher Scientific) containing 3% FBS and 20 mM HEPES. To the cell suspension, 20 μM coelenterazine-h or 3 μM furimazine was added to record luminescence spectra by PMA-12. The obtained fluorescence and luminescence spectra were used for the estimation of energy transfer efficiencies of the biosensors.

Komatsu N., Terai K., Imanishi A., Kamioka Y., Sumiyama K., Jin T., Okada Y., Nagai T, & Matsuda M. (2018). A platform of BRET-FRET hybrid biosensors for optogenetics, chemical screening, and in vivo imaging. Scientific Reports, 8, 8984.

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Chloride Imaging Using SuperClomeleon FRET Sensor

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Chloride imaging was carried out as previously described.²³ (link)^,²⁴ (link)^,⁹² (link) The change of chloride concentration was determined using the YFP/CFP FRET-based chloride sensor SuperClomeleon. Briefly, the excitation filter FF02-438/24 (Semrock) was used to excite the sensor and the emission filters FF01-483/32 and FF01-542/27 were used to capture CFP and YFP fluorescence, respectively, for 180 s. The fluorescence intensity of each channel of glial cell body was calculated by subtracting the background intensity. Then, the ratio YFP/CFP (R) was calculated for each frame accordingly. The average ratio of the 10 s (R₀) before stimulation was considered as basal level. The following YFP/CFP ratios were calculated by comparing the difference with the average basal level of the 10 s (R/R₀) before stimulation.

Wang L., Graziano B., Encalada N., Fernandez-Abascal J., Kaplan D.H, & Bianchi L. (2022). Glial regulators of ions and solutes required for specific chemosensory functions in Caenorhabditis elegans. iScience, 25(12), 105684.

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FRET-based Chloride Imaging Protocol

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For chloride imaging, the excitation filter FF02-438/24 (Semrock) was used to excite the sensor and the emission filters FF01-483/32 and FF01-542/27 were used to capture CFP and YFP fluorescence, respectively, for 120 seconds. SuperClomeleon is a YFP/CFP FRET-based chloride sensor in which YFP/CFP fluorescence ratio decreases upon binding to chloride ions (Park et al., 2021 ). For each frame, the fluorescence intensity of each channel in the region of interest was calculated by subtracting the background adjacent to the ROI. Consequently, the ratio (R) YFP/CFP was calculated. The average R of the 10 seconds before touch was considered R₀. R/R₀ was then plotted to visualize chloride changes during touch stimulation. The R/R₀ change (ΔR/R₀) was calculated as the difference between the average R/R₀ of the last 10 seconds of the recording (Fig. 3C-D during B) and the 10 seconds before touch (Fig. 3C-D during A).

Fernandez-Abascal J., Johnson C.K., Graziano B., Wang L., Encalada N, & Bianchi L. (2021). A glial ClC Cl− channel mediates nose touch responses in C. elegans. Neuron, 110(3), 470-485.e7.

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Rac1 FRET Biosensor Imaging

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We established a stable cell line of 3T3 cells expressing a Rac1-FRETbiosensor (Moshfegh et al., 2014) . For imaging, cells were plated on glass coverslips with crossbow micropatterns. After 4 hr of adhesion, cells were imaged by epifluorescence using a Luca R camera (Andor on an Olympus IX71 microscope with a 603 magnification objective; Olympus PlanApo 603, NA 1.45). The same excitation and dichroic mirrors (e.g., FF02-438/24, BS: FF-458-DiO2; Semrock) were used for the sequential acquisition of donor and acceptor images. A filter wheel was used to switch emission filters of donor (mCerulean, Em: FF01-483/32) and FRET acceptor (Em: FF01-542/ 27). Image processing included registration, flat-field correction, background subtraction, segmentation, and FRET/donor ratio calculations. FRET ratio images were then aligned and averaged as described in the Supplemental Information.

Remorino A., De Beco S., Cayrac F., Di Federico F., Cornilleau G., Gautreau A., Parrini M.C., Masson J.B., Dahan M, & Coppey M. (2017). Gradients of Rac1 Nanoclusters Support Spatial Patterns of Rac1 Signaling. Cell reports, 21(7).

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Monitoring ATP Dynamics in Arabidopsis

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Wild-type Arabidopsis and pgr5 mutants expressing recombinant Nano-lantern(ATP1) were grown in Murashige and Skoog medium, and the first or second leaves of 7-day-old plants were harvested. Detached leaves were injected with coelenterazine h as described above and then, to assess their luminescence, were immediately placed onto the stage of a microscope system (Ti-E (Nikon), which included a camera, iXon Ultra 897 (Andor Technology); objective lens, CFI Plan Apo VC 20× (NA = 0.75) (Nikon) and filters, FF02-438/24 (actinic light, Semrock), FF01-500/24 (mVenus excitation, Semrock) and FF01-542/27 (mVenus emission, Semrock).
The settings were as follows: exposure, 1 s; dead time, 300 ms; binning, 1; EMgain, Max. The luminescence in the leaves was measured in the presence and absence of actinic light ( max = 438 nm; 9.1 mW cm -2 ).
Rise and decay signals over a 7-s period were acquired immediately after dark-to-light or light-to-dark transitions and then were fit to single exponential curves. The luminescence rise and decay rates are reported herein as the half-life of each exponential curve.
We performed at least three replicates for each analysis.

Sato R., Kawashima R., Trinh M.D.L., Nakano M., Nagai T, & Masuda S. (2019). Significance of PGR5-dependent cyclic electron flow for optimizing the rate of ATP synthesis and consumption in Arabidopsis chloroplasts. Photosynthesis research, 139(1-3).

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