The pCMV- IRES-EGFP (pEGFP) plasmid was used to create the pEGFP-GVs complexes, which were then incubated with HEK293 cells for 6 h (pEGFP-GVs@HEK293 cells). The resulting pEGFP-GVs@HEK293 cells were then subjected to 1 min of acoustic irradiation at 0, 0.1, 0.2, or 0.5 MPa pressure, followed by observation under a fluorescence microscope (EVOS M7000, Thermo Fisher, USA) after 48 h or quantitative analysis of their transfection efficiencies by flow cytometry (CytoFLEX LX, Beckman, USA). To compare the gene transfection efficiencies mediated by intracellular cavitation and extracellular cavitation, pEGFP-GVs complexes were incubated with HEK293 cells, B1610 cells, MC38 cells, and C6 cells for 6 h, yielding pEGFP-GVs@HEK293 cells, pEGFP-GVs@B1610 cells, pEGFP-GVs@MC38 cells and pEGFP-GVs@C6 cells, respectively. For 1 min, these pEGFP-GVs@cells were irradiated at 0.5 MPa of acoustic pressure. In terms of extracellular cavitation, equivalent pEGFP-GVs complexes were added to these HEK HEK293 cells, B1610 cells, MC38 cells, and C6 cells and immediately treated with acoustic irradiation at 0.5 MPa for 1 min. These irradiated cells were examined under a fluorescence microscope (EVOS M7000, Thermo Fisher, USA) after 48 h and quantitatively analyzed using flow cytometry (CytoFLEX LX, Beckman, USA).
Evos m7000
Manufactured by Thermo Fisher Scientific
Sourced in United States
The EVOS M7000 is a compact, automated microscope designed for live-cell imaging. It features high-quality optics, precise environmental control, and intuitive software for capturing high-resolution images and time-lapse videos of cells in culture.
Automatically generated - may contain errors
Lab products found in correlation
Celleste image analysis software, by Thermo Fisher Scientific (4 mentions)
Hoechst 33342, by Thermo Fisher Scientific (4 mentions)
Upper transwell chamber, by Corning (3 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (3 mentions)
Cytoflex lx, by Beckman Coulter (2 mentions)
Hcs mitochondrial health kit, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 594, by Thermo Fisher Scientific (2 mentions)
Propidium iodide, by Thermo Fisher Scientific (2 mentions)
Slide 8 well, by Ibidi (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
48 well plate, by Corning (2 mentions)
Glass bottom 96 well plate, by Thermo Fisher Scientific (2 mentions)
Plo laminin, by Iwaki (2 mentions)
Fluoromount g mounting medium, by Southern Biotech (2 mentions)
Orca er ccd camera, by Hamamatsu Photonics (1 mentions)
Plan apochromat 1.4 na objective, by Zeiss (1 mentions)
Novaseq 6000, by Illumina (1 mentions)
Cryostar nx70, by Thermo Fisher Scientific (1 mentions)
Leukocyte acid phosphatase kit, by Merck Group (1 mentions)
Mitosox, by Thermo Fisher Scientific (1 mentions)
107 protocols using evos m7000
1
Acoustic Cavitation-Mediated Gene Transfection
2
Microscopy Techniques for Cell Imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Phase contrast imaging was performed using the EVOS M7000 imaging system (Invitrogen by Thermo Fisher Scientific). H&E histology slides were imaged using the EVOS M7000 imaging system (Invitrogen by Thermo Fisher Scientific). Trichrome histology slides were scanned by the CHOP pathology core and images captured using Image Scope. For differential interference contrast (DIC) imaging live cells were imaged with a Zeiss Axiovert 200M platform using a 40× EC Plan Neo Fluar, 1.3 N.A., or a 63× Plan Apochromat, 1.4 N.A. objective (Carl Zeiss, Jena, Germany) equipped with an Orca ER CCD camera (Hamamatsu, Bridgewater, NJ, USA). Images were captured using Slidebook 6 imaging software (Intelligent Imaging Innovations, Denver, CO, USA) using an Orca ER CCD camera (Hamamatsu).
3
Spatial Transcriptomic Profiling of Primary PN Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Optimal cutting temperature (OCT) compound embedded frozen primary PN samples were sectioned on a Cryostar NX70 cryostat (ThermoFisher) at 10 μm sections and mounted on spatial transcriptomic capture slides (Visium, 10 × Genomics). The optimal tissue section permeabilization time (18 min) was established using preliminary test sections. 4 PN sections were permeabilized and resulting capture sections underwent methanol fixation and were stained with H&E and imaged on an Evos M7000 (ThermoFisher) with brightfield settings. RNA libraries were generated and subsequently sequenced at 70,000 read pairs per spot (Novaseq6000, Illumina).
4
Immunofluorescence Staining of Baculovirus-Expressed Protein
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunofluorescence staining was visualized using a fluorescence microscope (EVOS M7000; Thermo Fisher Scientific), as described previously [32 (link)]. Sf9 cells were seeded into the wells of 24-well plates and infected with recombinant baculovirus at an MOI of 1. At 48 h after infection, cells were fixed with 4% paraformaldehyde for 20 min and permeabilized with 0.2% Triton X-100 for 20 min. After several washes with PBS, the samples were blocked with 5% bovine serum albumin at 37 °C for 30 min and incubated with gE-specific antibody (1B11, 1:2000) overnight at 4 °C. After three washes, the samples were incubated with Alexa Fluor 647-conjugated goat anti-mouse IgG secondary antibody (1:1000 dilution) at 37 °C for 1 h. Nuclei were counterstained with DAPI. Fluorescence was visualized using a fluorescence microscope and images were analyzed with Fiji (Image-J) software (NIH; Bethesda, MD, USA). Bac–WT cells were treated in the same manner.
5
Mitochondrial Function Assay of Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The mitochondrial functionality was analyzed by HCS Mitochondrial Health Kit (Thermo Fisher Scientific), following the manufacturer’s procedures. Briefly, hFM-MSCs and hAFSCs were treated with 0.1 μM of BPA, BPS, PFOS, and PFOA, alone or in combination, for 24 h. Then, cells were incubated with the MitoStain for 30 min at 37 °C and then fixed in paraformaldehyde 4% for 10 min. Nuclei were counterstained by Hoechst 33342. To quantify the fluorescence, 4 different fields for each sample were acquired by EVOS M7000 (Thermo Fisher Scientific) and analyzed by Celleste Image Analysis Software (Thermo Fisher Scientific).
6
Osteoclast Differentiation Assay with RLE
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RAW 264.7 cells were treated with 100 ng/mL RANKL and supplemented with different dosages of RLE for 7 days to determine the efficacy of RLE on osteoclast differentiation. Next, mature osteoclasts were washed with Dulbecco’s phosphate buffer saline (DPBS) and stained with TRAP using a commercial acid phosphatase leukocyte kit (Sigma, #387A). For cytochemical staining of TRAP-positive cells, deparaffinized tissue sections or 10% formaldehyde-fixed cells were stained for TRAP following the manufacturer’s protocol. TRAP-positive cells were counted microscopically (EVOS M7000, Thermo Fisher Scientific Inc., Waltham, MA, USA), and their activity was determined as described previously [20 (link)].
7
Mitochondrial ROS Imaging in Ovarian Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The fixed bone tissues were then washed with 1x phosphate-buffered saline (PBS) and incubated with 1x PBS containing 5 μM MitoSOX (Invitrogen, Waltham, MA, USA, #M36008) for 40 min at 37 °C in a dry oven. The specimens were washed with 1x PBS. The slides containing ovarian tissues were observed with a fluorescence microscope (EVOS M7000, Thermo Fisher Scientific, Waltham, MA, USA). All parts of each slide were observed, and representative images were captured and analyzed by the ImageJ program (National Institutes of Health, Bethesda, MD, USA).
8
Transwell Migration Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the transwell assay, after transfection for 24 h, cells suspended in serum-free DMEM medium were seeded into the upper transwell chamber(Corning). In the lower chamber, DMEM containing 20% FBS was filled in the well. After culturing for 36 hours, the filters were fixed in methanol and stained with 0.1% crystal violet. The upper cells of the filters were gently abraded, and the lower cells migrated across the filters were imaged and counted under the microscope(EVOS M7000; Thermo Fisher) at 100×. The numbers of migrated cells were counted and calculated by ImageJ (NIH).
9
Evaluation of TGF-β1 Paracrine Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cyb5r3-Scr and Cyr5r3-KD MLE12 cells were cultured as described above, until recombinant TGF-β1 or PBS was added to the cell culture medium at 10 ng/mL for 8 hours. Cells in all groups were then extensively washed with plain HITES media and cultured in fresh TGF-β1–free culture medium for 48 hours. After 48 hours, the media were collected, and the MLE12 cells were washed and pelleted for RNA extraction. This 48-hour conditioned medium was used to culture primary murine fibroblasts for 24 hours (in a 1:1 ratio, conditioned medium to fresh fibroblast culture media), after which stimulatory potential was assessed using light microscopy (EVOS M7000, Thermo Fisher Scientific) observation and by quantitative PCR analysis of the lysed fibroblasts.
10
Quantitative Cas12a-based NA Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An RPA mix was prepared by combining 240 nM of target‐specific forward and reverse primers, 0.8 U µL−1 RNase inhibitor, and target NA. Upon the addition of 14 mM MgOAc, the reaction proceeded at 37 °C for 20 min. The amplified target NAs were then mixed with ≈50 Cas12a/gRNA‐HMPs in 1× NEBuffer 2.1 containing the F‐Q probe (2 µM) and RNase inhibitor (0.8 U µL−1). After incubation (1 h, 37 °C), HMPs were introduced to the microfluidic device using manual pipettes or disposable eye droppers. After capturing individual HMPs on‐chip traps, the fluidic chamber was filled with HFE‐7500 oil. After an additional incubation period (30 min, 37 °C), HMPs were imaged using EVOS M7000 (ThermoFisher Scientific) and the image was analyzed with the customized ML algorithm. For a given NA target, the fluorescent signal was measured from about 15 target‐specific HMPs and the mean value was used as an analytical metric. The CLAMP signal was corrected by subtracting the background signal measured without target NA.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!