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The Thermo Fisher Scientific D1845 is a laboratory equipment product. It is designed to perform a core function within a laboratory setting. No further details are provided to maintain an unbiased and factual approach.

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Lab products found in correlation

Axio zoom v16, by Zeiss (4 mentions) Eclipse ti inverted microscope, by Nikon (1 mentions) D1818, by Thermo Fisher Scientific (1 mentions) Synergy 2, by Agilent Technologies (1 mentions) Oregon green 488 dextran, by Thermo Fisher Scientific (1 mentions) D1820, by Thermo Fisher Scientific (1 mentions)

9 protocols using d1845

1

Dextran Diffusion Assay for Endothelial Barrier

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The barrier function of the iEC vessel models was assessed by measuring dextran diffusion across the vessel endothelium as previously described[30 (link)]. A 1 μM solution of dextran (FITC-conjugated 40 kDa, D1845; TRITC-conjugated 70 kDa, D1818, both ThermoFisher Scientific) was prepared in iEC media. For each vessel, 5 μL of dextran solution was injected through the small lumen port. Dextran diffusion was imaged with the Nikon TI® Eclipse inverted microscope every 5 min over 15 min. Effective permeability coefficients (expressed in cm/s) were calculated using the following equation[31 (link)]:
P=1I0[IF−I0tF−t0](D4),
where Io is the total initial intensity outside the vessel, If is the total intensity outside the vessel at 15 min, to is the initial time point, tf is the final time point of 15 minutes, and D is vessel diameter.
Virumbrales-Muñoz M., Ayuso J.M., Loken J.R., Denecke K.M., Rehman S., Skala M.C., Abel E.J, & Beebe D.J. (2022). Microphysiological model of the renal cell carcinoma to inform anti-angiogenic therapy. Biomaterials, 283, 121454.
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2

Quantifying Blood-Brain Barrier Permeability

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Evans blue (EB) staining was performed to evaluate brain vascular leakage. After HH treatment, mice were injected via the tail vein with 4 mL/kg of 2% EB in saline. Approximately 1 h after circulation, the mice were anaesthetized and perfused with physiological saline solution (0.9%) to remove the intravascular dye. Half of the brain was immediately weighed, homogenized in 1 mL of 50% trichloroacetic acid solution, and then centrifuged at 15,000 × g for 30 min. The supernatant was then diluted 1:3 with ethanol, and its absorbance was measured at 630 nm using a microplate reader (Synergy 2™, Bio-Tek, US). The other half of the brain was fixed and sectioned for confocal imaging with a 647 nm laser by a Leica SP8. The mice were also intravenously injected with 40 kD FITC-dextran (D1845, Thermo; 4 mL/kg of 10 mg/mL in saline), which circulated for 15 min. At the end of the circulation period, the mice were promptly euthanized and perfused with physiological saline solution (0.9%) via the left ventricle to remove the blood.
Xue Y., Wang X., Wan B., Wang D., Li M., Cheng K., Luo Q., Wang D., Lu Y, & Zhu L. (2022). Caveolin-1 accelerates hypoxia-induced endothelial dysfunction in high-altitude cerebral edema. Cell Communication and Signaling : CCS, 20, 160.
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3

Choroid Plexus Organoid Permeability

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ChP permeability was determined by the influx of fluorescein isothiocyanate–dextran (FITC‐dextran) of 40 kDa (D1845; Thermo Fisher Scientific). Briefly, ChPOs were preincubated with the HSV‐1 virus (3000 pfu/organoid). Following the treatment, ChPOs were carefully washed twice with Hank's balanced salt solution (HBSS), and then dextran solution (140 μg/ml in HBSS) was added and incubated for 1 h. Next, the CSF‐like liquid was collected from the cystic structures of the ChPOs. The fluorescent intensity of the solution was determined using a multimode Spectramax M2 microplate reader, with the wavelengths of excitation and emission 494 and 518 nm, respectively. The permeability was determined as the amount of dextran that entered the CSF‐like liquid in 60 min.
Qiao H., Chiu Y., Liang X., Xia S., Ayrapetyan M., Liu S., He C., Song R., Zeng J., Deng X., Yuan W, & Zhao Z. (2023). Microglia innate immune response contributes to the antiviral defense and blood–CSF barrier function in human choroid plexus organoids during HSV‐1 infection. Journal of Medical Virology, 95(2), e28472.
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4

Automated Fluorescent Droplet Preparation

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1 μM solution of fluorescein isothiocyanate (FITC) labeled dextran (D1845, Thermo Fisher) was automatically loaded into a syringe and emulsified using the same protocol as ddPCR. Then, the FITC droplets were collected into a 96-well plate by the liquid handler. The robot exchanged the oil phase of FITC drops from 2% HFE to 5% FC40. The robotic arm transferred the well plate to the thermocycler, which closed and performed a dummy cycle (no heating or cycling). After the cycle, the thermocycler opened, and the robotic arm moved the well plate back to the pipettor region. The liquid handler performed another oil exchange to revert back to 2% HFE and moved the well plate to automated syringes.
Tran T.M., Kim S.C., Modavi C, & Abate A.R. (2022). Robotic automation of droplet microfluidics. Biomicrofluidics, 16(1), 014102.
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5

Visualizing Lymphatic Conduit Network

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Fluorescently labeled dextran was injected s.c. to aid in visualization of the conduit network. Various sizes of dextran-FITC (Oregon Green 488 Dextran, 10 kDa (Thermo Fisher Scientific, catalog D7170), dextran-FITC, 40 kDa (Thermo Fisher Scientific, catalog D1845), and rhodamine B–dextran, 70 kDa (Thermo Fisher Scientific, catalog D1841), doses (2.5 μg, 5 μg, and 10 μg), and time points (3, 5, 10, or 30 minutes) were assessed during preliminary studies (data not shown). Transit of 10 and 40 kDa dextran–FITC to LNs occurred within 2 minutes. A solution of 2.5 μg dextran-FITC (40 kDa or 10 kDa) in 20 μL PBS concomitant with harvesting at 5 minutes after injection provided a clear and distinguishable dextran-FITC fluorescence signal. The excised dLNs were placed in optimal cutting temperature (OCT) compound (Scigen Scientific, catalog 4583) on dry ice, and 6- or 40-μm cryosections were prepared for immunofluorescence microscopy.
Li L., Wu L., Kensiski A., Zhao J., Shirkey M.W., Song Y., Piao W., Zhang T., Mei Z., Gavzy S.J., Ma B., Saxena V., Lee Y.S., Xiong Y., Li X., Fan X., Abdi R, & Bromberg J.S. (2023). FRC transplantation restores lymph node conduit defects in laminin α4–deficient mice. JCI Insight, 8(8), e167816.
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6

Dextran Uptake Assay in Zebrafish

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The dextran uptake assay was adapted from previously described fluorescent dextran assays in zebrafish 28 . 40 kDa fluorescein conjugated dextran (D1845, Fisher Scientific) was dissolved in H2O as 50 mg/ml stock solution. To perform the dextran uptake assay, fluorescein-dextran stock solution was diluted with H2O to make 4 mg/ml solution, and 5 μl of working solution was injected intraperitoneally into adult fish. At 3 hours post injection, zebrafish tailfins were imaged under Zeiss AxioZoom V16 microscope with ZEN pro 2012. at magnification of 40X. For each fish, the widest blood vessels from 5 fin rays were measured and averaged to determine the fluorescence intensity using Fiji (v.2.3.0).
Sun F., Ou J., Shoffner A.R., Luan Y., Yang H., Song L., Safi A., Cao J., Yue F., Crawford G.E, & Poss K.D. (2022). Enhancer selection dictates gene expression responses in remote organs during tissue regeneration. Nature cell biology, 24(5), 685-696.
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7

Dextran Uptake Assay in Zebrafish

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The dextran uptake assay was adapted from previously described fluorescent dextran assays in zebrafish 28 . 40 kDa fluorescein conjugated dextran (D1845, Fisher Scientific) was dissolved in H2O as 50 mg/ml stock solution. To perform the dextran uptake assay, fluorescein-dextran stock solution was diluted with H2O to make 4 mg/ml solution, and 5 μl of working solution was injected intraperitoneally into adult fish. At 3 hours post injection, zebrafish tailfins were imaged under Zeiss AxioZoom V16 microscope with ZEN pro 2012. at magnification of 40X. For each fish, the widest blood vessels from 5 fin rays were measured and averaged to determine the fluorescence intensity using Fiji (v.2.3.0).
Sun F., Ou J., Shoffner A.R., Luan Y., Yang H., Song L., Safi A., Cao J., Yue F., Crawford G.E, & Poss K.D. (2022). Enhancer selection dictates gene expression responses in remote organs during tissue regeneration. Nature cell biology, 24(5), 685-696.
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8

Dextran Tracking in Mouse Uterus

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Mice were given tail vein injections of 200 μl (25 mg/mL) of FITC-conjugated 40 kDa dextran (Invitrogen D-1820) at E3.5 and 10 kDa dextran (Invitrogen D-1845) at E6.5 [41 (link)–43 (link)]. After 10 minutes, animals were euthanized. Uteri and implantation sites were dissected in cold phosphate buffered saline, fixed in Carnoy’s solution and embedded in paraffin wax. Sections (7 μm) were deparaffinized, rehydrated and mounted in Vectashield medium containing DAPI or stained for Notch1 prior to mounting. Dextran was administered to 3 mice at each stage. Specific staining was performed at least 3 times and 5 different uterine sections or implantation sites were analyzed at each stage.
Shawber C.J., Lin L., Gnarra M., Sauer M.V., Papaioannou V.E., Kitajewski J.K, & Douglas N.C. (2015). Vascular Notch proteins and Notch signaling in the peri-implantation mouse uterus. Vascular Cell, 7, 9.
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9

Dye Labeling of Zebrafish Embryos

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WT and zep mutant embryos were treated with 0.003% phenylthiourea (PTU; P7629) in E3 beginning at 24 hpf to reduce pigmentation. For microinjections, embryos were anesthetized in 0.02% tricaine and 40 kDa dextran-FITC molecules (Invitrogen D-1845) at a concentration of 5 mg/ml were injected into the circulation of 3 days post fertilization (dpf) zebrafish embryos. Embryos were transferred to fresh 0.003% PTU/E3 and kept in the dark at 28°C for 24 hours post injection, then imaged.
Kroeger PT J.r., Drummond B.E., Miceli R., McKernan M., Gerlach G.F., Marra A.N., Fox A., McCampbell K.K., Leshchiner I., Rodriguez-Mari A., BreMiller R., Thummel R., Davidson A.J., Postlethwait J., Goessling W, & Wingert R.A. (2017). The zebrafish kidney mutant zeppelin reveals that brca2/fancd1 is essential for pronephros development. Developmental biology, 428(1), 148-163.
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