Wound healing was monitored using a BioStation IM-Q Time Lapse Imaging System (Nikon) and 4T1 cell tracking was performed using Volocity® Software -Tracking in Volocity Software (Perkin Elmer). Cells were randomly selected for live cell tracking analyses and cell nucleus position identified in each frame in a 20 minute interval over 4 hours and subsequently connected by the built in tracking algorithm followed by quantification of average speed, distance and directionality, which was defined as displacement divided by the total distance, i.e. a cell moving in a straight line defined as 1.
Biostation im q time lapse imaging system
Manufactured by Nikon
Sourced in Japan
The BioStation IM-Q Time Lapse Imaging System is a laboratory equipment product manufactured by Nikon. It is designed to capture time-lapse images for biological research and observation. The system utilizes a high-resolution camera and motorized stage to acquire images at user-specified intervals over extended periods of time.
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5 protocols using biostation im q time lapse imaging system
1
Live Cell Tracking of Wound Healing
2
Time-Lapse Imaging of Cell Culture Dynamics
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Twenty thousand cells per quadrant were plated in a 35mm Hi‐Q4 culture dish (Ibidi, Planegg, Germany). ARN‐3236 was added, and time‐lapse phase‐contrast images (4 z‐sections, 2 μm each) were acquired every 2 min for a total of 24 h using a BioStation IM‐Q time‐lapse imaging system (Nikon, Tokyo, Japan) fitted with a temperature‐controlled growth chamber. Video analysis was done through nis‐elements viewer 4.50 imaging software (Nikon), and Imaris (Bitplane, Zürich, Switzerland) was used to export the video.
3
Time-Lapse Imaging of Mitosis in HeLa Cells
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HeLa‐H2B/tub cells 30 expressing H2B‐mCherry and EGFP‐α‐tubulin were kindly provided by Jan Ellenberg (EMBL Heidelberg, Germany) and used for time‐lapse microscopy as described previously 67 . Briefly, cells were plated on μ‐slide 8‐well ibiTreat chambers (Ibidi) and imaged at 5‐min intervals for 48 h on a Nikon BioStation IM‐Q Time Lapse Imaging System using a 20×/0.8 NA air objective, a 1.3 megapixel cooled monochrome camera (Nikon), and Nikon software for image acquisition. Alternatively, HeLa‐H2B/tub cells were imaged at 15‐min intervals on a DeltaVision deconvolution microscope using a 20×/0.75 air objective.
HeLadox‐YFP‐TIARr cells were plated on a 12‐well glass bottom plate (Cellvis) and imaged at 15‐min intervals for 12 h on a Nikon Ti‐E microscope with an integrated perfect focus system using a 40x/0.95 NA air objective, a 4.2 megapixel cooled monochrome sCMOS pco.pge camera, and NIS elements JOBS software for image acquisition. All cells were kept in 10% FBS/DMEM at 37°C and 5% CO2 during imaging. Image processing was performed using ImageJ software (NIH,http://rsbweb.nih.gov/ij/ ).
HeLadox‐YFP‐TIARr cells were plated on a 12‐well glass bottom plate (Cellvis) and imaged at 15‐min intervals for 12 h on a Nikon Ti‐E microscope with an integrated perfect focus system using a 40x/0.95 NA air objective, a 4.2 megapixel cooled monochrome sCMOS pco.pge camera, and NIS elements JOBS software for image acquisition. All cells were kept in 10% FBS/DMEM at 37°C and 5% CO2 during imaging. Image processing was performed using ImageJ software (NIH,
4
Quantifying Phagocytosis of Fluorescent Beads by MDMs
5
Sunitinib Exposure in HMEC-1 Cells
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HMEC-1 cells (2×104/500 µl) with sunitinib (6 µM) for 72 h or HMEC6µM cells (2×104/500 µl) without sunitinib (6 µM) for 72 h were incubated in 4-well Hi-Q4 dishes (Biovalley) at 37°C in an incubator with 100% humidified atmosphere at 5% CO2. The images were recorded in real time using the BioStation IM-Q Time Lapse Imaging system (Nikon Corporation). Following the addition of sunitinib, culture plates were immediately put into the culture chamber of the time-lapse imaging system. The video recording usually began after a lapse of 15 min, which was required for the choice of image fields under microscopy.
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