Adult flies of the respective genotype were collected and etherized. Pictures were taken with an ES120 camera (Optronics, Goleta, CA, USA) mounted to a Wild M5 stereomicroscope (Leica, Wetzlar, Germany). Dehydrated wings of female flies were embedded in Euparal (Carl Roth, Karlsruhe, Germany) and documented with an ES120 camera (Optronics, Goleta, CA, USA) coupled to a Zeiss Axiophot (Carl Zeiss, Jena, Germany). Pictures were recorded using Pixera viewfinder software version 2.0. Alternatively, uncoated etherized flies were pictured with a table top scanning electron microscope NeoScope (JCM-5000 SEM; Nikon, Tokyo, Japan) using proprietary software. The wing or eye area was determined using the freehand tool of Image J (open source). Only the SEM pictures of female flies were used for the quantification of micro- and macrochaetae, respectively. The numbers of bristles were determined as described before [57 ,58 (link),59 (link),60 (link)]. Statistical analysis was conducted by ANOVA two-tailed test for multiple comparisons using Tukey–Kramer’s or Dunnett’s approach, as indicated: *** p ≤ 0.001 highly significant; ** p ≤ 0.01 very significant; * p ≤ 0.05 significant; not significant (n.s.) p > 0.05. The figures were assembled using Corel Photo Paint, Corel Draw and BoxPlotR software.
Es120 camera
Manufactured by Optronics
Sourced in United States
The ES120 camera is a high-performance digital camera designed for laboratory and scientific applications. It features a 12-megapixel image sensor and is capable of capturing images with a resolution of 4,096 x 3,072 pixels. The camera is equipped with a C-mount lens interface, allowing for compatibility with a wide range of lenses.
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Lab products found in correlation
6 protocols using es120 camera
1
Fly Wing and Eye Imaging Protocol
2
Autophagy Induction in Drosophila Larvae
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For analyzing autophagy, third instar larvae were dissected in ice-cold PBS and stained with the nuclear dye Hoechst 33342 (1:2000) for thirty minutes at room temperature before the fat body was removed and incubated in the dark for two minutes with LysoTracker® (1:100 in PBS) (Invitrogen; Carlsbad CA, USA). The staining solution was removed, and the tissue was mounted in Vectashield (Vector lab; Burlingame CA, USA) and immediately documented with a Zeiss ApoTome using Axio-Imager Software (Carl Zeiss AG; Oberkochen, Germany). For amino acid deprivation, third instar larvae were starved on apple-juice plates [25% apple-juice, 2.4% saccharose, 2.4% agar] without any further food source for 14 hours. The larvae were dissected, and after fixation for 10 minutes in 4% paraformaldehyde the fat body was mounted in glycerol (80%) and directly examined with a Zeiss Axioskop coupled to an ES120 camera (Optronics, Goleta, CA), using Pixera (Santa Clara, CA) Viewfinder software, version 2.0. Pictures were assembled with Corel (Mountain View, CA) PhotoPaint and CorelDRAW software, version 9.0.
3
Detailed Wing Development Analysis
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Adult males, three to five days old, were used for analysis. Dehydrated wings were mounted in Euparal (Roth; Karlsruhe, Germany). Wing area was measured using Image J software (oval selection for total wing size in two measurements that were sampled). Cell number was determined by counting the individual trichoma on the wing blade in three defined 10.000 μm2 squares localized in the L4/L5 field next to the posterior cross vein. Subsequently, the total cell number for the determined wing area was calculated, as was cell size. Pictures were assembled using Corel Draw and Corel Photo Paint. Flies and wings were photographed with an ES120 camera (Optronics; Goleta CA, USA) using Pixera Viewfinder software, version 2.0.
To investigate developmental timing, offspring of six parallel inter se crosses with the genotype w1118; cycGHR7/+ was counted at days 9 to 20. Since the cycGHR7 mutant carries a mini-white+ gene [24 (link)], the homozygous cycGHR7 and the heterozygous cycGHR7/+ flies could be distinguished by red and orange eye color, respectively, from the white-eyed control siblings.
To investigate developmental timing, offspring of six parallel inter se crosses with the genotype w1118; cycGHR7/+ was counted at days 9 to 20. Since the cycGHR7 mutant carries a mini-white+ gene [24 (link)], the homozygous cycGHR7 and the heterozygous cycGHR7/+ flies could be distinguished by red and orange eye color, respectively, from the white-eyed control siblings.
4
Quantifying Cell Division in Wing Discs
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Cells in M phase within the posterior compartment were counted based on PH3 signals. The posterior compartment was determined by GFP labelling from en-Gal4-GFP [46 (link)]. Cell number was related to total size of the respective wing discs using ImageJ. p53R-GFP expression in salivary glands was examined by measuring signal intensity of 12 nuclei from seven different glands each (n = 84 nuclei), using the mean intensity of Pzg signals as internal standard. Wings from female flies were dehydrated in ethanol, mounted in Euparal (Roth, Karlsruhe, Germany) and documented with an ES120 camera (Optronics, Goleta CA, USA) connected to a Zeiss Axiophot (Carl Zeiss AG, Jena, Germany) using Pixera Viewfinder software, version 2.0. Female flies were etherized before taking pictures from the heads with an ES120 camera coupled to a Leica Wild M3C Stereomicroscope (Leica, Wetzlar, Germany). Size of female eyes (UASp-lacZ/+; ey-Gal4/+ and ey-Gal4/+; UAS-mei-41/+) or wings (UASp-lacZ/+; en-Gal4-GFP/+ and en-Gal4-GFP/+; UAS-mei-41/+) was measured using Image J. Statistical analysis was conducted by ANOVA using a two-tailed Tukey-Kramer or Dunnett’s test for multiple comparisons. ***p < 0.001 highly significant; **p < 0.01 very significant; *p < 0.05 significant; not significant (ns) p > 0.05. Box plots were compiled using the online plotting tool BoxPlotR (http://shiny.chemgrid.org/boxplotr/ ).
5
Dehydration and Imaging of Insect Wings
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Adult wings were dehydrated in ethanol and mounted in Euparal (Roth, Karlsruhe, Germany), pictures of wings were taken with an ES120 camera (Optronics, Goleta, USA) using Pixera viewfinder software version 2.0. Wing size was measured with ImageJ programme.
6
Analyzing Female Eye Size via Microscopy
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Adult eyes of females were documented with an ES120 camera (Optronics, Goleta CA, USA) using Pixera viewfinder software version 2.0. For quantification, eye size of five females each was measured with Image J and eye area was calculated. Statistical significance of probes was determined according to Student's T-test (http://www.physics.csbsju.edu/stats/t-test.html ) and p-value was scaled accordingly: p>0.05 (not significant, n.s.); p<0.05 (weakly significant; *); p<0.01 (significant; **); p<0.001 (highly significant; ***).
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