Fluorescent images were taken with a Leica M205 FA stereomicroscope using an EGFP filter [Leica Microsystems Inc., Concord, Ontario Canada] at 1, 2, 3, and 4 dpf to monitor the chemical phenotypes. Unhatched embryos were removed from their chorion manually. The zebrafish were then anesthetised with a 50/50 E2 media/100 ppm clove oil solution and embedded in 2.5% methylcellulose [Sigma-Aldrich, St. Louis, Missouri, USA], a viscous solution that allows the fish to stay in a desired orientation for imaging.
Egfp filter
Manufactured by Leica
Sourced in Canada, United States
The EGFP filter is a high-performance optical filter designed for the detection and imaging of enhanced green fluorescent protein (EGFP) in biological samples. It is optimized to selectively transmit the specific wavelength range associated with EGFP fluorescence, allowing for the efficient separation and visualization of EGFP-labeled specimens.
Automatically generated - may contain errors
3 protocols using egfp filter
1
Zebrafish Imaging with Fluorescence
2
In Vivo Lymphatic Remodeling Assay
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Skin samples from C57BL/6J Prox-1GFP mice were dissected from areas of aberrant lymphatics (equivalent areas used in sham control mice). Lymphatic vessels were visualized using Prox-1GFP epifluorescence under a fluorescence stereo-dissecting microscope with an eGFP filter (Leica Microsystems). Between 15 and 30 images were taken per mouse, blinded, and lymphatic channels measured for aperture in ImageJ. All image measurements were pooled per mouse to calculate average lymphatic widths.
BmL3 were washed before incubation with 50 μM Alexa Fluor 546 NHS ester (succinimidyl ester) (Thermo Fisher Scientific) in Fluorobrite DMEM (Thermo Fisher Scientific) for 2 hours. C57BL/6J Prox-1GFP transgenic mice were injected with 400 fluorescent BmL3 as described above. After 3 hours and 1–6 dpi in mice, areas of subcutaneous tissues where lymphatic remodeling occurs were imaged as above (DsRed and eGFP filters).
BmL3 were washed before incubation with 50 μM Alexa Fluor 546 NHS ester (succinimidyl ester) (Thermo Fisher Scientific) in Fluorobrite DMEM (Thermo Fisher Scientific) for 2 hours. C57BL/6J Prox-1GFP transgenic mice were injected with 400 fluorescent BmL3 as described above. After 3 hours and 1–6 dpi in mice, areas of subcutaneous tissues where lymphatic remodeling occurs were imaged as above (DsRed and eGFP filters).
3
Measuring Cell-to-Cell Dye Movement in Maize Leaves
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We modified the DANS dye-loading assay (Cui et al., 2015) to analyze cell-to-cell CF movement in maize leaves. The H 2 O 2 treatment was performed by spraying 50 mM H 2 O 2 on intact plants every hour for 3 h prior to the DANS assay. The adaxial surface of leaf five from 3-week-old maize plants was gently abraded with a 5-mm-wide strip of very fine grain sand paper (2000 grain) to remove the cuticle. To minimize dye loss to the xylem or phloem, a 1 ml droplet of CFDA solution (1 mM) was loaded between two major veins on the adaxial surface of the gently abraded region. After 5 min, the leaf segment was excised, and the dye movement was observed under a Leica M205 FA stereoscope with an eGFP filter (Leica Microsystems, USA). ImageJ was used to quantify the region containing the dye by counting the color pixels. PD permeability was quantified as the ratio of the fluorescent signal intensity on the abaxial surface divided by the signal intensity on the adaxial surface of the leaf.
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