For all experiments, 2 × 105 PC12 cells/well (American Type Culture Collection, Manassas, VA, USA) were seeded onto six-well culture dishes coated with 50 μg/ml collagen type I and 0.1% poly-L-lysine (70 to 150 kDa; Sigma-Aldrich). Cells were allowed to attach to culture surfaces for 24 hours in RPMI 1640 medium supplemented with 1% antibiotic/antimycotic solution, 5% FBS and 10% horse serum (all obtained from Gibco/Invitrogen). After cell attachment, control wells were changed to 0.1% serum RPMI 1640 medium supplemented with 50 ng/ml recombinant βNGF (BioShop Canada, Burlington, ON, Canada) or sterile water vehicle (control). The remaining wells were subjected to 1.5 ml of conditioned media collected from the experiments described above (SS and HMS AF cultures and SS and HMS NP cultures). Neurite outgrowth was monitored for 4 days. Three random phase images per sample were taken from each individual experiment (n = 3), and the neurites per cell body were counted and scored as either ≤3 or >3 neurites/cell. Phase images were captured using a Zeiss Axiovert 40C microscope (Carl Zeiss Microscopy, Thornwood, NY, USA) equipped with a Canon PowerShot A640 digital camera (Canon USA, Melville, NY, USA) attached to a Zeiss MC 80 DX 1.0× tube adapter (Carl Zeiss Microscopy). LIVE/DEAD assays were performed and quantified as described above.
Powershot a640 digital camera
Manufactured by Canon
Sourced in Japan, United States
The PowerShot A640 is a digital camera manufactured by Canon. It features a 4.0 megapixel image sensor and a 4x optical zoom lens.
Automatically generated - may contain errors
Lab products found in correlation
Axiovert 40c inverted microscope, by Zeiss (3 mentions)
Horse serum, by Thermo Fisher Scientific (1 mentions)
Antibiotic antimycotic solution, by Thermo Fisher Scientific (1 mentions)
Axiovert 40c microscope, by Zeiss (1 mentions)
Collagen type 1, by Merck Group (1 mentions)
Poly l lysine (pll), by Merck Group (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Pc12 cells, by American Type Culture Collection (1 mentions)
Eclipse 80i epifluorescence microscope, by Nikon (1 mentions)
Axioplan light microscope, by Zeiss (1 mentions)
Axionvision sample images software, by Zeiss (1 mentions)
Cary eclipse, by Agilent Technologies (1 mentions)
Easysep human cd45 depletion kit, by STEMCELL (1 mentions)
Axiovert 40 cfl microscope, by Canon (1 mentions)
Interleukin 3 (il 3), by Thermo Fisher Scientific (1 mentions)
Stem cell factor (scf), by Thermo Fisher Scientific (1 mentions)
Thrombopoietin tpo, by Thermo Fisher Scientific (1 mentions)
Myelocult h5100, by STEMCELL (1 mentions)
Pituitary extract, by Merck Group (1 mentions)
Nerve growth factor (ngf), by Merck Group (1 mentions)
9 protocols using powershot a640 digital camera
1
Neurite Outgrowth Evaluation in PC12 Cells
2
Wound Healing Assay in HUVECs
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Wound healing assays were performed as previously described.29 (link) HUVECs were allowed to grow to full confluence in six-well plates. Subsequently, cells were wounded by pipette tips and washed twice with media to remove detached cells, and photomicrographs of initial wounds were taken using Canon PowerShot A640 camera (at ×100 magnification). Thereafter, RPMI 1640 containing 1% FBS was added to prevent proliferation. At the time of wounding, cells were treated with DMSO or 1 μg/mL–10 μg/mL dose of BPC-157. The wound area was photographed immediately after wounding (0 hours), and again at 12 hours postscratch using brightfield exposure at ×10 magnification on a Nikon eclipse 80i epifluorescence microscope. The images were captured using a Canon PowerShot A640 digital camera. The distance between the edges of the wound were measured at ten different areas from the wound edge to edge using Spot software. The measurements were then converted into a percentage using the following formula: % of wound remaining = (measurement at time 12 hours/measurement at time 0 hours) ×100; then the percentage of wound closure was calculated as follows: 100% – % of wound remaining.
3
Fixation, Clearing, and Microscopic Examination of Nematodes
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The nematodes were fixed in A.F.A. (70° GL ethanol, 93%; formaldehyde, 5%; glacial acetic acid, 2%) at 70 °C, for 48 h, transferred to a solution containing 70% ethanol and 5% glycerin, cleared and mounted on slides with lactophenol (one part distillated water, two parts glycerin, one part lactic acid, one part phenic acid) and observed under a light microscope.
Measurements were performed to the nearest micron (mean ± S.D. (range)) and were conducted on three mature adult specimens and 10 embryonated eggs in utero. Measurements were conducted with an Axioplan Zeiss light microscope (Carl Zeiss, Germany) equipped with a Canon Power-Shot A640 digital camera (Canon, China) and Zeiss AxionVision Sample Images Software (Carl Zeiss, Germany) for image analysis. Specimens deposited in the Harold W. Manter Parasite Collection at the University of Nebraska-Lincoln (UNL/USA), registration number HWML 67092, were examined for comparative purposes. Representative specimens were deposited in the Helminthological Collection of the Oswaldo Cruz Institute (Rio de Janeiro, Brazil) under the registration number CHIOC 38941.
Measurements were performed to the nearest micron (mean ± S.D. (range)) and were conducted on three mature adult specimens and 10 embryonated eggs in utero. Measurements were conducted with an Axioplan Zeiss light microscope (Carl Zeiss, Germany) equipped with a Canon Power-Shot A640 digital camera (Canon, China) and Zeiss AxionVision Sample Images Software (Carl Zeiss, Germany) for image analysis. Specimens deposited in the Harold W. Manter Parasite Collection at the University of Nebraska-Lincoln (UNL/USA), registration number HWML 67092, were examined for comparative purposes. Representative specimens were deposited in the Helminthological Collection of the Oswaldo Cruz Institute (Rio de Janeiro, Brazil) under the registration number CHIOC 38941.
4
Sensitive Detection of G-quadruplex DNA
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The one-tube SDA/GQ-RCA reaction mixture (25 μL) contained an aliquot of t-RCA products (2.5 μL of 25 μL preceding RCAP), 500 μM dNTPs, 100 nM of the 2nd dumbbell padlock DNA (DP2), 2.5 U KF, 10 U Nb.BbvCI, 115 U phi29 DNA polymerase, and 15 μM ThT in a reaction buffer (50 mM potassium acetate, 20 mM Tris-acetate, 10 mM magnesium acetate, 100 μg/ml BSA, and 2 mM KCl; pH 7.9). The reaction mixture was incubated at 32 °C for 30 min, and the reaction was terminated by heating at 95 °C for 10 min. To visualize the enhanced ThT fluorescence due to ThT intercalation into the G-quadruplex, the heat-quenched reaction products in a transparent PCR tube, after cooling at 8 °C for 5 min, were photographed under UV illumination using a PowerShot A640 digital camera (Canon Inc., Tokyo, Japan). The fluorescence intensity of each reaction sample (25 μL) containing ThT intercalated into the G-quadruplex, which was diluted with the same volume of double-distilled water, was measured using a fluorescence spectrophotometer (Cary Eclipse, Agilent Technologies, Santa Clara, CA, USA); λex = 425 nm and λem = 488 nm; PMT voltage = 600.
5
ADSC Wound Healing Assay
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ADSCs were plated at a density of 8 × 104 cells/well in six-well plates. After the cells had reached 70% confluence, wounds were produced by scratching with 200 μL pipette tips. The medium was replaced with or without PMA (100 nM), and the cells were incubated for up to 24 h (1 day). Images were captured using an Axiovert 40 °C inverted microscope (Carl Zeiss, Germany) equipped with a Powershot A640 digital camera (Canon, Japan) at 4 h and 24 h.
6
Long-Term Culture of CD34+ Cells
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LTC assays were performed by plating 1 × 103 CD34+ BM cells in duplicate on irradiated MS-5 murine stromal cells and/or autologous/allogeneic MSCs using Myelocult H5100 (StemCell Technologies, Vancouver, BC, Canada) in the presence of cytokines (20 ng/ml SCF, 20 ng/ml IL-3 and 20 ng/ml thrombopoietin (TPO) from PeproTech, London, UK). After 4 weeks, hCD45+ cells were isolated using EasySep Human CD45 Depletion Kit (StemCell Technologies, cat. no. 18259). Isolated CD45+ cells were washed in phosphate-buffered saline, counted and then plated into methylcellulose as mentioned below. The rest of the cells were harvested and stored as cell pellet for future use for genomic and single-nucleotide polymorphism assays. Images of LTC were acquired with a Zeiss (Cambridge, UK) AxioVert 40 CFL microscope using 5 × lens and connected to Canon (Melville, NY, USA) PowerShot A640 digital camera.
7
Culturing and Resting Corneal Endothelial Cells
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CECs were first cultured in a previously reported medium (MitoM) [9 (link)] containing OptiMEM-I supplemented with 8% FBS, 20 ng/ml of nerve growth factor (NGF; Sigma-Aldrich Co.), 5 ng/ml of epidermal growth factor (EGF; Sigma-Aldrich Co.), 100 µg/ml of pituitary extract (Sigma-Aldrich Co.), 200 µg/l of calcium chloride (Sigma-Aldrich Co.), 20 µg/ml of ascorbic acid (Sigma-Aldrich Co.), 0.08% chondroitin sulfate (Sigma-Aldrich Co.), and antibiotics. Isolated cells were incubated at 37 °C in a 5% CO2 humidified atmosphere. The medium was changed every third day until 80% confluence. At passage 1, the CECs were subcultured at a 1:2 split ratio. Population 1 continued to be cultured in MitoM, while population 2 was cultured in OptiMEM-I supplemented with only 8% FBS and 1% antibiotics (RestM) up to 80% confluence for an additional passage. We refer to the RestM procedure as “resting” because it lacks growth factors to decrease proliferation rates. An Axiovert 40 CFL contrast microscope (CFL; Carl Zeiss AG, Oberkochen, Germany) featuring a PowerShot A640 digital camera (Canon Inc., Tokyo, Japan) was used to register cell morphology.
8
VSMC Wound Healing Assay
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VSMCs were plated at a density of 2.5×105 cells/well in six-well plates. After the cells reached 80% confluence, the cells were serum-starved in 0.5% FBS-containing medium for 24 h. Then, the cells were wounded with 200 μl pipette tips, and the starting point was marked on the bottom of each plate. The medium was exchanged for 0.5% FBS (serum-deprived medium) or for 10% FBS (serum-containing medium) medium, and the cells were incubated for 0, 12, and 24 h. Images were captured using an Axiovert 40C inverted microscope (Carl Zeiss, Thornwood, NY, USA) equipped with a PowerShot A640 digital camera (Canon, Osaka, Japan).
9
Assessing Cell Migration using Scratch Assay
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SMCs (1 × 105 cells) were plated onto 6-well plates. After the cells reached 90% confluence, the cells were serum-starved for 4 h and then treated with mitomycin (5 μg/mL) for 1 h. The cells were scratched with 200 μL pipette tips, and the starting point was marked with a marker pen at the bottom of the plate. The cells were washed with PBS three times, the medium was replaced with 0.5% FBS medium containing flagellin (100 ng/mL), and the cells were incubated for 24 h. Images were captured using an Axiovert 40 C inverted microscope (Carl Zeiss, Oberkochen, Germany) equipped with a PowerShot A640 digital camera (Canon, Tokyo, Japan).
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