We used Canon Rebel T3i (Canon Inc. (Ōta, Tokyo, Japan)) to capture macroscopic images and then stitched them into timelapse movies using Persecond for Mac version 1.5 (Flixel Inc. (Toronto, Canada)). For all the timelapses reported in our study, we used a remote control to automatically capture an image every 4 min and published the movie files at 16 fps.
Rebel t3i
Manufactured by Canon
Sourced in United States, Japan
The Canon Rebel T3i is a digital single-lens reflex (DSLR) camera. It features an 18-megapixel CMOS sensor, DIGIC 4 image processor, and a 3-inch vari-angle LCD screen. The camera can record full HD 1080p video at 30 frames per second. It is compatible with a wide range of Canon EF and EF-S lenses.
Automatically generated - may contain errors
Lab products found in correlation
Axio zoom v16, by Zeiss (4 mentions)
Axiocam mrc, by Zeiss (2 mentions)
Rebel t3i camera, by Canon (1 mentions)
Rotor hda, by Singer Instruments (1 mentions)
Szx16, by Olympus (1 mentions)
Mp e 65 mm 1 5 macro lens, by Canon (1 mentions)
Alexa fluor 488 tagged goat anti rabbit secondary antibody, by Jackson ImmunoResearch (1 mentions)
Eclipse ni u, by Nikon (1 mentions)
Deltavision deconvolution microscope, by Cytiva (1 mentions)
Stemi 2000 c, by Zeiss (1 mentions)
Axiocam mrm, by Zeiss (1 mentions)
Axiovert 200m microscope, by Zeiss (1 mentions)
Mp e 65 mm 1 5x macro lens, by Canon (1 mentions)
Axiocam 506, by Zeiss (1 mentions)
Bz 9000, by Keyence (1 mentions)
Ix70 microscope, by Olympus (1 mentions)
Trolox, by Merck Group (1 mentions)
Rattler plating beads, by Zymo Research (1 mentions)
29 protocols using rebel t3i
1
Timelapse Image Capture and Stitching
2
Imaging Methods for Specimen Documentation
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The single known specimen is preserved as part and counterpart and required special imaging methods for viewing and documentation. A broad habitus view of the entire specimen was provided by combining separate images into a composite form. Each component image was taken from separate focal planes that subsequently were integrated by CombineZM® from a series of images. These different images then were stitched with Adobe Photoshop® CS 3 software under cross-polarized light with a Canon Rebel T3i camera equipped with a MP-E 65 mm lens and a MT 24 EX Canon Twin Flash. For comparison of the specimen to earlier studies that used standard light-microscopy, fully reflected vertical light as well as directed side light from different angles was used. In order to document specimen relief, two techniques were applied. The first approach was stereo-imaging that employed the Canon Rebel T3i camera. A second procedure involved virtual surface reconstruction resulting from stacks of images recorded on a Zeiss Axiophot microscope attached to a Skopetech DCM 510 camera. Cross-polarized light was provided by multiple, optical-fiber light sources. Virtual surfaces were calculated in Image Analyzer® software.
3
High-Throughput Screening of E. coli Mutants
4
High-throughput Screening of Bacterial Mutants
5
Photomorphogenic Apical Hook Analysis
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Seedlings were grown in a light-sealed box equipped with an infrared light source (880 nm LED) and a spectrum-enhanced camera (EOS035 Canon Rebel T3i) modified by Hutech technologies with a built-in, clear, wideband-multicoated filter. The camera was operated by EOS utility software. For light exposure experiments, 40 hours after germination (hooks in maintenance phase) the box was opened and a light source (4 µmoles/m2/s) was placed in front of the plate. Angles between the cotyledons and the hypocotyl axis were measured every 3 hours in dark and every hour during light exposure until complete opening using ImageJ (http://rsb.info.nih.gov/ij/ ) software. The outer angle of the apical hook is reported in the graphs. Representative experiments are shown. At least 3 independent experiments were performed and sample size is indicated in the figure legends.
6
High-throughput Screening of Bacterial Fitness
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For each strain probed, a single source plate was generated and transferred to the final screening plate using a pinning robot (Biomatrix 6). Chemical perturbations (1% SDS; 0.25 mM and 0.5 mM EDTA; 0.25% SDS and 0.25 mM EDTA; pH 4.8; pH 8.2; 0 mM NaCl; 600 mM NaCl; 50 µg/mL vancomycin) were added to the LB agar or adjusted before dispensing into the plates. MMT buffer (1:2:2 molar ratio of d ,l -malic acid, MES, and Tris base) was added to the media adjusted to pH 4.8 and 30 mM HEPES-KOH buffer to pH 8.2. On each screening assay plate, the parental strain, the single mutants, and the double mutants were arrayed, each in approximately 20 replicates per plate. The plates were incubated at 37°C (or for three conditions at 18°C, 25°C, and 42°C) for ~18 h and imaged under controlled lighting conditions (S&P Robotics) using an 18-megapixel Canon Rebel T3i (Canon). Colony size of endpoint images was quantified as fitness readout using the image analysis software Iris. Fitness ratio and statistical analysis were carried out by ChemGAPP as described below.
7
Angle Measurement of Cotyledon Closure
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Seedlings were grown in a light-protected box equipped with an infrared light source (880 nm LED) and a spectrum-enhanced camera (EOS035 Canon Rebel T3i) modified by Hutech technologies with a built-in, clear, wideband-multicoated filter. The camera was operated by EOS utility software. Angles between the cotyledons and the hypocotyl axis were measured every 3 h in the dark until opening using ImageJ (http://rsb.info.nih.gov/ij/ ) software. The complementary angle of the measured angle is reported in the graphs (180° represents full closure and 0° full opening). More information can be found in ref. 1 .
8
Genetic Interaction Assay Protocol
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Genetic interaction assay was performed as described in Banzhaf et al., 2020 (link). For each probed strain, a single source plate was generated and transferred to the genetic interaction plate using a pinning robot (Biomatrix 6). On each genetic interaction assay plate, the parental strain, the single deletion A, the single deletion B and the double deletion AB were arrayed, each in 96 copies per plate. Genetic interaction plates were incubated at 37°C for 12 hr and imaged under controlled lighting conditions (spImager S and P Robotics) using an 18-megapixel Canon Rebel T3i (Canon). Colony integral opacity as fitness readout was quantified using the image analysis software Iris (Kritikos et al., 2017 (link)). Fitness ratios were calculated for all mutants by dividing their fitness values by the respective WT fitness value. The product of single mutant fitness ratios (expected) was compared to the double mutant fitness ratio (observed) across replicates. The probability that the two means (expected and observed) are equal across replicates is obtained by a Student's two‐sample t‐test.
9
Photomorphogenic Apical Hook Analysis
10
High-throughput Screening of Bacterial Mutants
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The E. coli KEIO collection 29 was arrayed on solid agar plates in 1536 format, and plates were incubated at room temperature for 30 hours. Medium used was LB 0% NaCl supplemented with 40 µg/ml Congo red (CR) and 20 µg/ml Coomassie blue.
The S. Typhimurium single gene deletion library 55 (link) was arrayed on solid agar plates in 384 format, and plates were incubated at room temperature for 48 hours. Medium composition was the same as for E. coli.
The P. aeruginosa PA14 single deletion transposon library 30 (link) was arrayed on solid agar plates in 384 format, and plates were incubated at room temperature. Medium used was TB (Tryptone Broth) supplemented with 20 µg/ml CR and 10 µg/ml Coomassie blue.
All plates were imaged under controlled lighting conditions (spImager S&P Robotics Inc.) using a 18 megapixel Canon Rebel T3i (Canon Inc. USA).
The S. Typhimurium single gene deletion library 55 (link) was arrayed on solid agar plates in 384 format, and plates were incubated at room temperature for 48 hours. Medium composition was the same as for E. coli.
The P. aeruginosa PA14 single deletion transposon library 30 (link) was arrayed on solid agar plates in 384 format, and plates were incubated at room temperature. Medium used was TB (Tryptone Broth) supplemented with 20 µg/ml CR and 10 µg/ml Coomassie blue.
All plates were imaged under controlled lighting conditions (spImager S&P Robotics Inc.) using a 18 megapixel Canon Rebel T3i (Canon Inc. USA).
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