After transferring H1299 cells and culturing them for 24 h, a straight wound was created using a 200-µL sterile tip in the middle of the well. The cells were washed with PBS twice to smooth the edges of the scratch and remove floating cells. After incubation (37°C, 5% CO2) for 0, 6, 12, and 24 h, images of the migrating cells were observed under an optical inverted microscope (Olympus Corp., Japan). The experiments were repeated 3 times.
Inverted optical microscope
Manufactured by Olympus
Sourced in Japan, United States
The Inverted optical microscope is a type of microscope that has the objectives and light source positioned below the specimen, allowing for the observation of samples from the underside. This design permits the use of specialised cell culture techniques and provides a clear view of the specimen.
Automatically generated - may contain errors
Lab products found in correlation
Matrigel, by BD (8 mentions)
Crystal violet, by Beyotime (3 mentions)
Dexamethasone (dex), by Merck Group (3 mentions)
Paraformaldehyde (pfa), by Merck Group (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Paraformaldehyde (pfa), by Boster Bio (2 mentions)
Oil red o, by Merck Group (2 mentions)
Inverted fluorescence microscope, by Olympus (2 mentions)
α mem, by Merck Group (2 mentions)
Ascorbic acid, by Merck Group (2 mentions)
β glycerophosphate, by Merck Group (2 mentions)
Alizarin red, by Merck Group (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Paraformaldehyde (pfa), by Beyotime (2 mentions)
Microplate reader, by Thermo Fisher Scientific (2 mentions)
Ultra low attachment six well plate, by Corning (1 mentions)
Transwell cell culture insert, by BD (1 mentions)
Uorescence microscope, by Zeiss (1 mentions)
Hoechst, by Beyotime (1 mentions)
91 protocols using inverted optical microscope
1
Cell Migration Scratch Assay
2
Visualizing Apoptosis and Pyroptosis
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To examine the morphological changes in apoptotic or pyroptotic cells, HK2 cells were seeded in six-well plates and subjected to the indicated treatments. Bright field images were captured using an optical inverted microscope (Olympus, Japan). All image data represent at least three random fields of view.
3
CRC Spheroid Formation and Analysis
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SNU-C5 CRC cells treated with free Dox or PrPC-Apt DOA were cultured in ultra-low attachment six-well plates (Corning, Corning, NY, USA) for spheroid formation. The CRC cells were incubated in RPMI 1640 medium and grown at 37 °C in a 5% CO2-humidified atmosphere. Spheroids were grown for 14 d and observed using an optical inverted microscope (Olympus, Tokyo, Japan).
4
Matrigel-Coated Transwell Invasion Assay
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Transwell cell culture inserts (BD Biosciences, Franklin Lakes, NJ), pre-coated with 8% Matrigel (BD Biosciences), were used to measure cell invasion. In brief, 200 μL cell suspension containing 1 × 104 OS cells (pretreated with 10 µg/ml mitomycin C for 30 min) was placed into the upper chamber, and 700 μL DMEM medium containing 10% FBS in the lower chamber. Then, the Transwell plates were cultured in 37 ℃ for 48 h. Thereafter, the upper chambers were removed and immersed in 4% paraformaldehyde (Boster) for 20 min to fix the cells. Then, cells were stained with 1% crystal violet solution for 15 min. After washing twice with PBS and scrubbing away noninvasive cells, the chamber conditions were recorded using an optical inverted microscope (Olympus; magnification times 100 ×).
5
Morphological Analysis of Apoptotic Cells
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To examine the morphological changes of apoptotic or secondary pyroptotic cells, HK2 cells were seeded in the 6-well plates and subjected to indicated treatments. Bright eld cell images were captured using the Optical inverted microscope (Olympus, Japan).All the image data displayed represents at least three random eld of view.
HK2 cell Hoechst/PI uorescent staining HK2 cells were seeded in the 24-well plates and treated with TNF-α and CHX. Then, cells were stained with Hoechst (Beyotime, China) and PI(BD, USA).Images were collected with a uorescence microscope(Carl Zeiss, Germany).
HK2 cell Hoechst/PI uorescent staining HK2 cells were seeded in the 24-well plates and treated with TNF-α and CHX. Then, cells were stained with Hoechst (Beyotime, China) and PI(BD, USA).Images were collected with a uorescence microscope(Carl Zeiss, Germany).
6
Mycobacterium bovis BCG Infection Model
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Bacterial strain, parasites, mice and infections. Mycobacterium bovis BCG, Pasteur strain were grown in Middlebrook 7H9 medium (Difco Laboratories; BD Biosciences, Franklin Lakes, NJ, USA) with 10% albumin dextrose catalase (ADC) (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) and 0.05% Tween 80. Cryopreserved blood pre-treated with Plasmodium yoelii (P. yoelii 17XNL) was defrosted and used to infect BALB/c mice. A total of 60 female BALB/c mice (6-week-old, 18-23 g) were purchased from Vital River Laboratory Technology Animal Co., Ltd. Histopathological analysis. Following sacrifice, histological examinations were performed. The livers were removed, fixed in 10% formaldehyde solution at 4˚C for 1 week, and then dehydrated. The paraffin sections (5 µm) were cut and hematoxylin and eosin (H&E) staining was performed at room temperature for 2 h. The results of the staining (magnification, x200) were analyzed with an optical inverted microscope (Olympus Corporation, Tokyo, Japan). Simultaneously, the number of granulomas in the liver was determined in the H&E-stained sections (5 sections per mouse, 3 mice per group).
7
Wound Healing Assay for Cell Migration
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The wound healing assay was conducted to assess the migratory capacity of cells. Initially, cells were seeded at a density of 4 × 105 cells per well in a six-well plate and maintained in serum-deprived medium until reaching approximately 90% confluence. Subsequently, the confluent cell monolayer was subjected to controlled mechanical injury by gently scraping with a sterile 1 mL pipette tip. Following injury, the cells were rinsed twice with sterile PBS. The remaining adherent cells were then subjected to OGD/R treatment and cultured under serum-deprived conditions for a duration of 48 hours. Photomicrographs capturing the wound area were acquired at both the initial time point (0 hours) and after the 24-hour incubation period, utilizing an inverted optical microscope manufactured by Olympus Corporation.
8
Adipogenic Differentiation of Mesenchymal Stem Cells
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Mesenchymal stem cells were seeded in six-well plates and cultured in basal medium for 24 h. Then, cells were cultured for 14 days with adipogenic medium, containing 0.5 mM isobutylmethylxanthine (MP Biomedicals, Santa Ana, CA, USA), 10 μg/ml insulin, 0.5 mM dexamethasone (MP Biomedicals, Santa Ana, CA, USA) and 60 mM indomethacin (MP Biomedicals, Santa Ana, CA, USA). The medium was refreshed every 3 days. The cells washed twice with PBS to remove medium and then fixed with 4% paraformaldehyde (Sigma-Aldrich, St. Louis, MO, USA). Then cells were washed gently three times with ddH2O and stained with Oil Red O (Sigma-Aldrich, St. Louis, MO, USA). Photographs were taken by the inverted optical microscope (Olympus, Tokyo, Japan).
9
Transwell Assay for Invasive Potential
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To evaluate the invasive potential of cells, a transwell assay was employed cells at a quantity of 4 × 105 cells were introduced into the upper chambers of transwell inserts featuring an 8 μm pore size (Millipore, USA). The cells were then cultivated in serum-deprived medium, while the lower chambers were supplied with complete medium supplemented with 15% fetal bovine serum (FBS). Following exposure to the OGD/R for a duration of 48 hours, the cells that had migrated to the lower surface of the upper chamber were fixed using a solution of 4% paraformaldehyde for 30 minutes. Following fixation, these cells were stained with 0.1% crystal violet (#C0121, Beyotime, China) for 15 minutes. Post-staining, the cells were washed twice with PBS, and the migrated cells were visualized and quantified through utilization of an inverted optical microscope manufactured by Olympus Corporation. The quantification process involved tallying the number of migrated cells, which facilitated subsequent data analysis.
10
Analyzing Trichoderma reesei Growth
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To analyze T. reesei vegetative growth, equal amounts of mycelia were inoculated on MM agar plates containing different carbon sources (glucose, glycerol, lactose, xylose, arabinose or Avicel) with or without exogenously adding copper and incubated at 30°C. To analyze T. reesei biomass on liquid MA medium containing 1% glucose, equal amounts of pre-cultured mycelia were inoculated and the mycelia collected at growth intervals were dried and weighed. For conidiation analysis, mycelia were inoculated on malt extract agar plates with or without 20 μM copper and incubated at 30°C for 5 days. The number of conidia was counted with a hemocytometer on an inverted optical microscope (Olympus, Tokyo, Japan).
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