Photographs of flies were taken with a binocular equipped with Leica DC480 digital camera using Leica IM50 Image Manager Software. We took photos of pairs of flies corresponding to each of the four comparisons (10 pairs for each comparison). Using ImageJ, we decomposed each picture in hue, saturation and brightness and measured hue mean pixel intensity in thorax + abdomen of each fly. We then calculated the hue difference between the darkest and the lightest fly for each pair (Table S9 -S12 ).
Dc480
Manufactured by Leica
Sourced in Canada
The DC480 is a digital camera designed for microscopy and documentation applications. It features a high-resolution sensor and high-quality optics to capture detailed images of samples under a microscope.
Automatically generated - may contain errors
Lab products found in correlation
Im50 image manager software, by Leica (5 mentions)
Macro apotome, by Zeiss (2 mentions)
Micro apotome, by Zeiss (1 mentions)
Dm4000b microscope, by Leica (1 mentions)
Bx41 fluorescence microscope, by Yokogawa (1 mentions)
Coolsnaphq2, by Universal Imaging (1 mentions)
Metaview software, by Universal Imaging (1 mentions)
Isopentane, by Merck Group (1 mentions)
Neo mount, by Merck Group (1 mentions)
Sucrose, by Merck Group (1 mentions)
10 protocols using dc480
1
Measuring Fly Hue Differences
2
Quantifying Drosophila Trident Pigmentation
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Flies were immersed in 75% Ethanol and pinned through the abdomen on a silicon substrate. Their thoraces were imaged with a binocular equipped with Leica DC480 digital camera using the Leica IM50 Image Manager software. An annular lamp was used to ensure homogeneous lighting. All pictures were taken with identical settings during the same session. To quantify the intensity of the trident, we delimited the trident using a polygon based on the suture anterior to the scutellum and the base of each dorsocentral macrochaetes using imagej (Rasband, 2016 ) (see Supporting information Figure S7 ). We measured the mean intensity of the pixels included in this polygon with the function “measure” of imagej . The obtained values were subtracted from 255 to get values comprised between 0 (white) and 255 (black). The trident of ten males was quantified for each tMSE haplotype.
3
Quantifying Aortic Plaque Deposition
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Lipid accumulation in the mouse aortic arch and the brachiocephalic artery was determined as previously described (13 (link)). In brief, the aortic arch and its branches were cut open and pinned to a flat surface. Samples were stained using Sudan IV and micrographs of the aortas were captured using a microscope (Leica MZ6) and camera (Leica DC480) and stained area measured by a blinded evaluator. The stained plaque area was subsequently measured by a second blinded evaluator using ImageJ software (NIH). Plaque coverage was calculated as the percentage of plaque area of the total surface area of the aortic arch with the aortic intercostal arteries denoting a caudal end of the arch and not including branching vessels. Plaque coverage was measured similarly in the brachiocephalic artery.
4
Imaging Abdominal Epidermal Cuticles and in situ Hybridization
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Adult cuticles and abdominal in situ hybridizations were imaged with a binocular equipped with Leica DC480 digital camera using the Leica IM50 Image Manager software. An annular lamp was used to ensure homogeneous lighting. Care was taken to use identical settings in each set of experiments. The higher magnification pictures showing y expression associated with bristles as well as abdominal epidermes of y-body-nEGFP pharates were acquired using a micro-apotome (Zeiss). nEGFP was measured in hemi-tergites A5, A6 and A7 using maximum intensity projections of stacks of 14 to 16 pictures (39 to 45 μm thick) using the Zen software.
5
Quantifying Abdominal Pigmentation and nEGFP Intensity
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Adult cuticles and abdominal in situ hybridizations were imaged with a binocular equipped with a Leica DC480 digital camera using the Leica IM50 Image Manager software. They were imaged using identical settings and an annular lamp to ensure homogeneous lighting. To quantify pigmentation, each entire hemi-segment was circled by hand. For A5 and A6, the melanic line at the dorsal limit of each hemi-segment (i.e the dorsal midline) separates the two hemi-segments. Cuticle pigmentation in hemi-tergites A5, A6 or A7 was measured as mean grey value using ImageJ. This value was subtracted from 255 to get a final pigmentation value comprised between 0 (white) and 255 (black).
Abdominal epidermes of t_MSE-nEGFP and ebony-nEGFP females were imaged using a Macro-Apotome (Zeiss). nEGFP intensity was measured in hemi-tergites A5, A6 or A7 using ImageJ in Maximum Intensity projections of 40 picture stacks.
Abdominal epidermes of t_MSE-nEGFP and ebony-nEGFP females were imaged using a Macro-Apotome (Zeiss). nEGFP intensity was measured in hemi-tergites A5, A6 or A7 using ImageJ in Maximum Intensity projections of 40 picture stacks.
6
Quantification of Nuclear HIF1α Immunofluorescence
7
Visualizing Drosophila Epidermal Gene Expression
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Cuticles were imaged as described in35 (link). Abdominal epidermes of females expressing nEGFP under the control of t_MSE, yellow and ebony regulatory sequences were imaged using a macro-apotome (Zeiss). Young adult females were imaged for t_MSE-nEGFP and ebony-nEGFP and female pupae for yellow-nEGFP as these genes are not expressed at the same developmental stage. The mean intensity of yellow-nEGFP was measured using ImageJ. Larvae showing H2B-GFP under the control of pnr-Gal4 and yellow-Gal4 were imaged with a binocular equipped with a Leica DC480 digital camera using the Leica IM50 Image Manager software. Pharates showing H2B-GFP under the control of pnr-Gal4 and yellow-Gal4 were imaged using an Olympus BX41 fluorescence microscope (objective 4 X) equipped with a Yokogawa spinning disc and a CoolSnapHQ2 camera controlled by Metaview software (Universal Imaging).
8
Visualizing Odontogenic Development in Mouse Incisors
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To visualize the early odontogenic areas in the mouse lower incisor region, Shh expression was detected using WISH in 93 Eda−/− and 57 Eda+/+ mouse embryos. Lower jaws were dissected at ED12.5–15.5, washed in RNase free PBS (pH 7.4) and fixed in 4% paraformaldehyde (PFA) solution overnight at 4°C. Specimens were hybridized according to a standard protocol. The probe for Shh was generated by in vitro transcription from cDNA fragment (kind gift from Dr. A. McMahon, Harvard University, Cambridge, Massachusetts). The hybridized samples were documented using a stereomicroscope Leica MZ6 connected with a digital camera Leica DC480 (Leica Microsystems GmbH, Wetzlar, Germany) and histologically processed.
Hybridized mouse embryonic heads were embedded in a series of graded solutions of sucrose (Sigma) diluted in PBS (pH 7.4). Then the specimens were embedded in O.C.T. Tissue Tek (Sakura) diluted 1:1 with 20% sucrose, frozen in isopentane (Sigma) cooled on dry ice to -60°C, and sectioned frontally on a cryostat microtome Mikrom HM 560 (Mikrom Walldorf, Germany) in series of 10 μm sections. The sections were post-fixed in 4% paraformaldehyde and counterstained by Nuclear Fast Red (Fluka), dehydrated, and mounted in Neomount (Merck).
Hybridized mouse embryonic heads were embedded in a series of graded solutions of sucrose (Sigma) diluted in PBS (pH 7.4). Then the specimens were embedded in O.C.T. Tissue Tek (Sakura) diluted 1:1 with 20% sucrose, frozen in isopentane (Sigma) cooled on dry ice to -60°C, and sectioned frontally on a cryostat microtome Mikrom HM 560 (Mikrom Walldorf, Germany) in series of 10 μm sections. The sections were post-fixed in 4% paraformaldehyde and counterstained by Nuclear Fast Red (Fluka), dehydrated, and mounted in Neomount (Merck).
9
Intestinal Morphometric Analysis
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The collected jejunum samples were fixed in formalin, then processed as previously described by Oladokun et al. (2021) (link). To access the villus height (VH), villus width (VW) and crypt depths (CD), measurements were made from the base of the intestinal mucosa to the villus tip, excluding the intestinal crypt for the VH, halfway between the base and the tip for the VW, and from the base upward to the region of transition between the crypt and villi for the CD. The ratio of VH to CD was also determined by measuring the total mucosa thickness. Ten measurements of each component per slide using an image processing and analysis system were carried out (Leica DC480; Leica Microsystems Imaging Solutions Ltd., Concord, ON, Canada).
10
Intestinal Morphological Analysis in Poultry
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The intestinal tissues were harvested from 12 birds per treatment group on d 14, and 24 birds per treatment group on d 28. All samples were processed using the same procedure as described by Oladokun et al. (2021) (link). After sample collection, the intestinal tissues were fixed, embedded in paraffin, sliced into sections (0.5 μm thick), and then stained with hematoxylin and eosin for morphological analysis. The small intestinal morphology slides were visualized using microscope and processed using Leica application (Leica DC480; Leica Microsystems Imaging Solutions Ltd., Concord, ON, Canada). Upon examination, the villus height was measured first; from the base of the intestinal mucosa to the tip of the villus excluding the crypt, then we measured the villus width (halfway between the base and the tip), and crypt depth was measured from the base upward to the region of transition between the crypt and villi. The villus height:crypt depth was then calculated. To ensure the reliability of data, ten morphometric measurements were evaluated per slide by the same individual. On d 28, 960 villi (10 villi/slide) were analyzed per section (duodenum, jejunum, and ileum) while 480 (10 villi/slide) villi were analyzed on d 14. Villi and crypts were selected based on their clear visibility and well-oriented positioning.
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